The present invention relates to a chemical compound. In particular the present invention relates to a chemical compound suitable for use as an anti-viral agent. The present invention also relates to the therapeutic use of the present chemical compound, to a pharmaceutical composition containing the present compound and to use of the present compound in the manufacture of a medicament.
Since the recognition of human acquired immunodeficiency syndrome (AIDS) much interest and research activity has been directed to its understanding and to attempting to provide a means of treatment. The human immunodeficiency virus (HIV) has been identified as the presumed aetological agent in AIDS. A large literature now exists related to the use of a wide variety of chemical compounds having as their object a demonstration of anti-viral activity with respect to HIV, hepatitus B virus (HBV), herpes and other viruses.
A class of compounds which has demonstrated anti-viral activity and which has been the subject of a large amount of research are nucleoside analogues.
An example of such a compound is xe2x80x9cAbacavirxe2x80x9d which is a substituted adenine analogue (Foster R. H. and Faulds D. Drugs 1998 55 729-736). This compound has entered clinical use due to the potential activity and stability of the compound displayed in preliminary work.
PCT/GB96100580 relates to a class of nucleoside analogues said to be highly active with respect to HIV. In particular PCT/GB96/00580 addresses the problem of providing compounds which are said to be highly potent in vitro viral inhibitors in both TKxe2x88x92 and TK+ cells. The compounds disclosed in PCT/GB96/00580 are phosphoramidates of purine or pyrimidine nucleoside analogues. Such compounds can however display chemical, for example acid, or biological, for example nucleoside phosphorylase, instability towards glycoside bond cleavage. Consequential deactivation may limit their potential clinical efficacy.
A compound however to be potentially useful in a clinical setting needs to exhibit a number of other properties as well as demonstrating, at least in in vitro tests, a sufficient and desired anti-viral activity. Primarily, these other properties comprise good pharmacokinetic properties, sufficient stability in the compound to permit its ease of handling and supply, and sufficiently low toxicity to permit its administration with an acceptable level of side effects to a patient in need of treatment for the viral infection in question.
In practice however it is frequently found that attempts to modify a compound demonstrating anti-viral activity in vitro, in order to improve its other properties, can have a detrimental effect on the anti-viral activity it displays. Ideally moreover any compound proposed for clinical trials needs also to have a ready means of administration and to be prepareable by an economically viable route.
It is an object of the present invention to provide a novel class of compounds exhibiting potent anti-viral, in particular anti HIV and/or HBV activity, in combination with good pharmacokinetic and stability properties and exhibiting sufficiently low toxicity so as to provide a compound having beneficial properties for clinical use.
According to the present invention there is provided a compound according to the following formula (1): 
wherein
Ar is an aryl group,
R1 and R2 are independently selected from the group comprising H, alkyl and aryl groups;
X is selected from the group comprising O, NH, NR4 and S wherein R4 is selected from the group comprising alkyl and aryl groups;
R5 is selected from the group comprising H, alkyl and aryl groups, wherein when R1 and R5 are each alkyl they may be linked to form a 5- or 6-membered ring;
and R3 is selected from the group comprising H, al, aryl, heterocyclic and polycyslic groups,
or a pharmaceutically acceptable derivative or metabolite thereof.
The present invention includes salts and physiologically functional derivatives of the presently defined compounds.
Reference in the present specification to an alkyl group means a branched or unbranched, cyclic or acyclic, saturated or unsaturated (e.g. alkenyl or alkynyl) hydrocarbyl radical. Where cyclic, the alkyl group is preferably C3 to C12, more preferably C5 to C10, more preferably C5 to C7. Where acyclic, the alkyl group is preferably C1 to C16, more preferably C1 to C6, more preferably methyl or ethyl.
Reference in the present specification to an aryl group means an aromatic group, such as phenyl or naphthyl, or a heteroaromatic group containing one or more, preferably one, heteroatom for example O, N and/or S, such as pyridyl, pyrrolyl, furanyl and thiophenyl. Preferably, the aryl group comprises phenyl or substituted phenyl.
The alkyl and aryl groups may be substituted or unsubstituted, preferably unsubstituted. Where substituted, there will generally be 1 to 3 substituents present, preferably 1 substituent. Substituents may include halogen atoms and halomethyl groups such as CF3 and CCl3; oxygen containing groups such as oxo, hydroxy, carboxy, carboxyalkyl, alkoxy, alkoyl, alkoyloxy, aryloxy, aryloyl and aryloyloxy; nitrogen containing groups such as amino, alkylamino, dialkylamino, cyano, azide and nitro; sulphur containing groups such as thiol, alkylthiol, sulphonyl and sulphoxide, heterocyclic groups which may themselves be substituted; alkyl groups, which may themselves be substituted; and aryl groups, which may themselves be substituted, such as phenyl and substituted phenyl. Alkyl includes substituted and unsubstituted benzyl. Reference in the present specification to alkoxy and aryloxy groups means alkyl-Oxe2x80x94and aryl-Oxe2x80x94 groups, respectively. Reference to alkoyl and aryloyl groups means alkyl-COxe2x80x94and aryl-COxe2x80x94, respectively.
Reference in the present specification to heterocyclic groups means groups containing one or more, optionally bridged, rings containing 1 to 6 heteroatoms in total. Each ring in the group may contain 3 to 12, preferably 1 to 6, atoms in total. At least one ring present contains 1 to 2 heteroatoms. Where two or more rings are present they may be fused or unfused. The rings can contain unsaturation. Heteroatoms includes 0, S and N. Examples of such heterocyclic groups containing one or more pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, yrazolidinyl, tetrahydrofuranyl, pyranyl, pyronyl, pyridyl, pyrazinyl, pyridazinyl, piperidyl, piperazinyl, morpholinyl, thionaphthyl, benzofuiranyl, isobenzofiyl, indolyl, oxyindolyl, isoindolyl, indazolyl, indolinyl, 7-azaindolyl, isoindazolyl, benzopyranyl, coumarinyl, isocoumarinyl, quinolyl, isoquinolyl, napthridinyl, cinnolinyl, quinazolinyl, pyridopyridyl, benzoxazinyl, quinoxadinyl, chromenyl, chromanyl, isochromanyl and carbolinyl.
References in the present specification to polycyclic groups means a group comprising two or more non-aromatic carbocyclic or heterocyclic rings which may themselves be substituted. Preferably the group contains 2 to 4 fused or non-fuised rings, each ring suitably containing 3 to 12 atoms, more suitably 4 to 10, more suitably 5 to 7, and even more suitably 5 to 6 atoms. The definitions of cyclic alkyl and heterocyclic rings given above also apply to the rings in the polycyclic groups.
Reference in the present specification to halogen means a fluorine, chlorine, bromine or iodine radical, preferably fluorine or chlorine radical.
The group Ar comprises a substituted or unsubstituted aryl group, wherein the term xe2x80x9caryl groupxe2x80x9d and the possible substitution of said group is as defined above. Preferably, Ar is a substituted or unsubstituted phenyl group. Particularly preferred substituents are electron withdrawing groups such as halogen (preferably chlorine or fluorine), trihalomethyl (preferably trifluoromethyl), cyano and nitro groups. Preferably, Ar is phenyl, 3,5-dichloro-phenyl, p-trifluoromethyl-phenyl, p-cyano-phenyl, or p-nitro-phenyl.
R3 is selected from hydrogen, alkyl, aryl, heterocyclic and polycyclic groups.
Preferably, R3 is a substituted or unsubstituted alkyl group. Preferably, R3 is a substituted or unsubstituted C1-6 alkyl group, more preferably an ethyl or methyl group.
Preferably, at least one of R1 and R2 is hydrogen. It will be appreciated that if R1 and R2 are different, the carbon atom to which they are bonded is an asymmetric centre. Preferably this carbon atom is chiral. When this carbon atom is chiral, the stereohemistry at this site may be D or L or mixed, with L-stereochemistry being preferred. R5 and R1 can be linked to form an alkylene bridge comprising 3 to 4 carbon atoms so as to form a 5- or 6-membered ring. Preferably R5 is hydrogen.
It will be appreciated that the group xe2x80x94NHxe2x80x94CHR1xe2x80x94CO2R3 corresponds to a carboxy-protected xcex1-amino acid. Preferably, the group R1 corresponds to the side chain of a naturally occurring amino acid such as Alanine, Arginine, Asparagine, Aspartic Acid, Cysteine, Cystine, Glycine, Glutanic Acid, Glutamine, Histidine, Isoleucine, Leucine, Lysine, Methionine, Phenylalanine, Proline, Serine, Threonine, Tryptophan, Tyrosine, Valine. Preferably, R1 is Me or PhCH2 corresponding to the side chain of alanine or phenylalanine, respectively. Preferably, the stereochemistry at the asymmetrc centre xe2x80x94CHR1xe2x80x94 corresponds to an L-amino acid.
It is a feature of the aryl ester phosphate compounds of the present invention that they exhibit significantly enhanced anti-viral efficacy in in vitro tests, in comparison to their corresponding nucleoside analogue, (1S,4R)-4-[2-anino-6-(cyclopropylamino)-9H-purin-9-yl]-2-cyclopentene-1-methanol, which is known as Abacavir and which has the following structural formula:

According to a further aspect of the present invention there is provided a compound of formula (II): 
wherein R1, R2, R3, R5 and X are as defined above, or a pharmaceutically acceptable derivative or metabolite thereof. Preferably X is O.
The intracellular generation of such anti-viral metabolites is an important feature of the invention for several reasons. In cases where the nucleoside is not a good substrate for host nucleotide kinases, activation will be poor and anti-viral efficacy low, even if the triphosphate is an excellent RT inhibitor. In such cases, the generation of the present metabolites may lead to a very significant enhancement in anti-viral action.
By xe2x80x9ca pharmaceutically acceptable derivativesxe2x80x9d is meant any pharmaceutically acceptable salt, ester or salt of such ester or any other compound which upon administration to a recipient is capable of providing (directly or indirectly) a compound of the present formula or present metabolite. Preferred xe2x80x9cpharmaceutically acceptable derivativesxe2x80x9d include sodium, succinate, fumarate, glutarate and D-tartrate salts.
By xe2x80x9cpharmaceutically acceptable metabolitexe2x80x9d is meant a metabolite or residue of a compound of the present formula or present metabolite which gives rise to reverse transcriptase inhibition exhibited by the present compounds.
According to a further aspect of the present invention there is provided a compound according to the present invention for use in a method of treatment, preferably in the prophylaxis or treatment of viral infection.
According to a further aspect of the present invention there is provided use of a compound according to the present invention in the manufacture of a medicament for the prophylaxis or treatment of viral infection.
According to a further aspect of the present invention there is provided a method of prophylaxis or treatment of viral infection comprising administration to a patient in need of such treatment an effective dose of a compound according to the present invention.
The viral infection may comprise any viral infection such as HIV and herpes vinis, including HSV I and HSV 2, CMV, VZV, EBV, HAV, HBV, HCV, HDV, HHV6, HHV7, HHV8, papilloma, adenoviruses, rabies and influenza.
Preferably, the viral infection comprises HIV or HBV infection, more preferably HIV-I or HIV-II. It is a feature of the present invention that the compounds exhibit good activity against HIV-I and HIV-II, and HBV.
According to a further aspect of the present invention there is provided a pharmaceutical composition comprising a compound of the present invention in combination with a pharmaceutically acceptable excipient.
According to a further aspect of the present invention there is provided a method of preparing a pharmaceutical composition comprising the step of combining a compound of the present invention with a pharmaceutically acceptable excipient.
Compounds of the present invention can demonstrate significant stability towards acid-mediated hydrolytic decomposition. The present compounds can thus be particularly suitable for oral administration under typical dosing conditions in humans as they can retain stability under the highly acidic environment of the stomach.
As the purine in compounds of formula (I) is a weak base (pKa=5.0) and the compounds of formula (a) demonstrate stability to acids, salts can be formed of compounds of formula (1) with acids, such as carboxlic acids and dicarboxlic acids. Such salts can be stable, crstalline solids, which can be beneficial in terms of improved shelf-life and ease of handling during manufacture into pharmaceutical compositions. Preferred carboxylic and dicarboxylic acids include malonic, succinic, glutaric, fumaric and tartaric acids. In contrast to the salts of compounds of formula (I), the free bases of compounds of formula (I) can be in a non-crystalline amorphous form which can be hygroscopic.
The P-OH group of compounds of formula (II) is a weak acid and can therefore form monobasic salts with bases to give, for example, sodium, potassium, ammonium, and triethylanmmonium salts. In. compounds of fonnula (II) when X is OH, dibasic salts can be formed. Such dibasic salts can be in the form of stable solids, which can provide benefits of improved shelf-life and ease of handling during manufacture into pharmaceutical compositions.
Compounds of the present invention can also demonstrate enhanced stability in biological media, for example, in human plasma. The increased half-life of compounds embodying the present invention in media such as human plasma may permit a pharamacokinetic advantage in dosing in humans in need of treatment.
The medicament employed in the present invention can be administered by oral or parenteral routes, including intravenous, intramuscular, intraperitoneal, subcutaneous, transdermal, airway (aerosol), rectal, vaginal and topical (including buccal and sublingual) administration.
For oral administration, the compounds of the invention will generally be provided in the form of tablets or capsules, as a powder or granules, or as an aqueous solution or suspension.
Tablets for oral use may include the active ingredients mixed with pharmaceutically acceptable excipients such as inert diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavouring agents, colouing agents and preservatives. Suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose, while corn starch and alginic acid are suitable disintegrating agents. Binding agents may include starch and gelatin, while the lubricating agent, if present, will generally be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate, to delay absorption in the gastrointestinal tract.
Capsules for oral use include hard gelatin capsules in which the active ingredient is mixed with a solid diluent, and soft gelatin capsules wherein the active ingredients is mixed with water or an oil such as peanut oil, liquid paraffin or olive oil.
Formulations for rectal administration may be presented as a suppository with a suitable base comprising for example cocoa butter or a salicylate.
Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known in the art to be appropriate.
For intramuscular, intraperitoneal, subcutaneous and intravenous use, the compounds of the invention will generally be provided in sterile aqueous solutions or suspensions, buffered to an appropriate pH and isotonicity. Suitable aqueous vehicles include Ringer""s solution and isotonic sodium chloride. Aqueous suspensions according to the invention may include suspending agents such as cellulose derivatives, sodium alginate, polyvinyl-pyrrolidone and gum tragacanth, and a wetting agent such as lecithin. Suitable preservatives for aqueous suspensions include ethyl and n-propyl p-hydroxybenzoate.
The compounds of the invention may also be presented as liposome formulations.
In general a suitable dose will be in the range of 0.01 to 10 mg per kilogram body weight of the recipient per day, preferably in the range of 0.2 to 1.0 mg per kilogram body weight per day. The desired dose is preferably presented once daily, but may be dosed as two, three, four, five or six or more sub-doses administered at appropriate intervals throughout the day. These sub-doses may be administered in unit dosage forms, for example, containing 10 to 1500 mg, preferably 20 to 1000 mg, and most preferably 50 to 700 mg of active ingredient per unit dosage form.
According to a further aspect of the present invention there is provided a process for the preparation of the present compound comprising reaction of a compound of formula 
with a compound of formula 
wherein R1, R2, R3, R5 and X have the meanings given above.
The reaction may be carried out under dry conditions at ambient temperature in tetrahydrofuran in the presence of N-methylimidazole, or by using t-butyl magnesium chloride and an excess of the appropriate phosphorochlorideate reagent.
Compounds embodying the present invention wherein Ar is replaced by H may be prepared from the acid form by treatment of the ester with an aqueous base.
Compounds wherein X is NH or NR4 can be prepared by treating the acid form (Xxe2x95x90O and R3=H) with amine.
The above starting material, (1S, 4R)-4-[2-amino-6-(cyclopropylamino)-9H-purin-9-yl]-2-cyclopentenel-methanol, is known as Abacavir and may be made by any procedure known in the art, for example by procedures described in European Patent Specification Number 0434450, PCT Patent Application No. PCT/GB95/02014, and PCT Patent Application No. PCT/EP98/02835, all of which are incorporated by reference thereto.
The invention will now be described with reference to the following Examples. It will be appreciated that what follows is by way of example only and that modifications to detail may be made whilst still falling within the scope of the invention.
The following anhydrous solvents and reagents were bought dry from Aldrich with sure seal stoppers: Dichloromethane (DCM), diethyl ether (Et2O), tetrahydrofuran (A), N-methyl imidazole (NMI), methanol (MeOH), dimethylformamide (DMF), pyridine (pyr), dioxane, and tBuMgCl. Triethylamine (NEt3) was dried by refluxing over CaH2 for several hours and then distilled off for immediate use.
Thin layer chromatography (tic) was performed on commercially available Merck Kieselgel 60F254 plates and the separated components were visualised using ultra violet light (254 nm and 366 nm), or by treatment with a 5% ethanolic solution of dodeca-molybdo-phosphoric acid (MPA) followed by heating. Column chromatography was performed using Woelm silica (32-63 mm) as the stationary phase.
All NMR spectral data, unless otherwise stated, were obtained in CDCl3. Proton and Carbon-13 nuclear magnetic resonance were recorded on a Bruker Avance DPX300 spectrometer with operating frequencies of 300 MHz and 75 MHz respectively. Phosphorous-31 NMR spectra were recorded on a Bruker Avance DPX300 spectrometer operating at 121 MHz, and are reported in units of xcex4 relative to 85% phosphoric acid as the external standard, positive shifts are downfield. The following abbreviations are used in the assignment of NMR signals: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), bs (broad signal), dd (double of doublets), dt (double of triplets).
Low resolution mass spectra were run on a VG Platform II Fisons instrument (Fisons, Altrincham, UK) (atmospheric pressure ionization, electrospray mass spectrometry) in either negative or positive ion mode.
High Performance Liquid Chromatography (HPLC) was performed on an SSODS2 reverse phase column with an eluent of water/acetonitrile. 100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins), with a flow rate of I ml/min and detection by UV at 254 mn. Standards: acetone (tR 4.54 mins), toluene (tR 10.21 mins). Final products showed purities  greater than 99%, with undetectable amounts of the parent nucleoside.
IUPAC nomenclature is used where possible, but for ease some compounds are abbreviated. Numbering is by conventional nucleoside numbering. 
For practical purposes, standard procedures are given where applicable.
To a stirring solution of anhydrous alcohol (10 mol eq) was added thionyl chloride (2 mol eq) dropwise at 0xc2x0 C., and the resulting solution stirred for Ihr. Upon rising to room temperature, the appropriate amino acid (1 mol eq) was added and the reaction heated at reflux for 6-16 hrs. Removal of the solvent and recrystallisation from methanol:ether gave the amino ester hydrochloride salts.
The appropriate amino acid (1 mol eq), para-toluene sulfonic acid (1.1 mol eq) and the appropriate alcohol (Imol eq) were heated under reflux in toluene (100 ml), under Dean and Stark conditions, for 6-16 hrs. On cooling to room temperature the solvent was removed under reduced pressure to give an oil. This was solubilised in dichloromethane (50 ml) and washed with 10% K2CO3 (50 ml), and water (50 ml), filtered and the filtratee reduced to dryness to give an oil. This was solubilised in the minimum amount of acetone and neutralised with 2M HCl, and then lyophilised to give the amino acid ester hydrochloride salts.
Phenyl dichlorophosphate (1 mol eq) and the appropriate amino acid ester hydrochloride salt (1 mol eq) were suspended in anhydrous dichloromethane (30-60 ml). Anhydrous triethylamine (2 mol eq) in anhydrous dichloromethane (30 ml) was added dropwise at xe2x88x9280xc2x0 C., and the reaction left to rise to room temperature overnight. The solvent was removed under reduced pressure, and under nitrogen, to give white solids. This was washed with anhydrous ether (2xc3x9725 ml), filtered and the filtrate reduced to dryness to give the products as crude oils. These were stored in anhydrous THF and used without any further purification.
(1 S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (1 mol eq) was dried by azeotroping with anhydrous pyridine (3xc3x975 ml), and then suspended in anhydrous THF (5-30 ml). To the suspension was added tBuMgCl (1-2 mol eq, 1.0M solution in THF) dropwise, and the resulting suspension stirred for 10 mins. The phosphorochloridate species (3 mol eq, solution in THF) was then added dropwise and the resulting solution stirred at room temperature for 24-96 hrs. The reaction was then quenched by the addition of sat.NC4Cl (0.1 ml), and after 10 mins the solvent was removed under reduced pressure. The crude product was purified by silica column chromatography.
L-Alanine methyl ester hydrochloride.
C4H10O2N1Cl1, MW=139.38.
This was synthesised according to Standard Procedure 1, using anhydrous methanol (34 ml, 0.84 mol), thionyl chloride (8.2 ml, 0.112 mol) and L-alanine (5.0 g, 0.056 mol). The product was isolated as a white solid (2.87 g, 36.7%).
1H NMR (D2O): xcex44.074.00 (1H,q,CH,J=7.22 Hz), 3.83 (3H,s,OCH3), 1.39-1.37 (3H,t,CH3).
13C NMR (D2O): xcex4171.5 (CO), 53.9 (O CH3), 49.1 (CH), 15.4 (CH3).
Phenyl-(methoxy-L-alaninyl)-phosphorochloridate.
C10H13O4N1Cl1P1, MW=277.65.
This was synthesised according to Standard Procedure 3, using L-Alanine methyl ester hydrochloride (2.0 g, 14.34 mmol), phenyl phosphorodichloridate (3.02 g, 2.14 ml, 14.34 mmol) and anhydrous triethylamine (2.90 g, 4.0 ml, 28.68 mmol). The product (3.91 g, 98.2%) was isolated as a colourless crude oil which was stored in anhydrous THF (40 ml) to give a 0.47M solution.
31p NMR: xcex49.28, 8.97 (1:1).
1H NMR: xcex47.39-7.34 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.29-7.20 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 4.494.37 (1H,q,NHala), 4.27-4.09 (1H,m,CHala), 3.78 (3H,d,OCH3), 1.52-1.49 (3H,dd,CH3).
13C NMR: 173.6 (CO), 150.1 (xe2x80x98ipsoxe2x80x99-Ph), 130.25 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 120.9 (xe2x80x98oxe2x80x99-Ph), 53.2 (OCH3), 51.0 (CH), 20.9 (CH3ala).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl-(methoxy-L-alaninyl)]-phosphate. Cf1490.
C24H30O5N7P1, MW=527.53.
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (500 mg, 1.75 mmol), tBuMgCl (1.0M solution in THF) (1.75 ml, 1.75 mmol) and phenyl-(methoxy-L-alaninyl)-phosphorochloridate (0.47M solution in THF) (1 1.17 ml, 5.24 mmol), in THF (30 ml) and stirring at room temperature for 70 hrs. The crude product was purified by column chromatography eluting with 3% MeOH in DCM and then 2% MeOH in DCM to give the product as a white foam (442 mg, 48%).
31P NMR (MeOH-d4): xcex43.97, 3.88.
1H N: xcex47.41 (1H,d,C8), 7.24-7.19 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.13-7.03 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 6.08 (1H,bs,NH), 5.98 (1H,q,H2xe2x80x2), 5.78 (t,H3xe2x80x2), 5.44 (1H,t,H1xe2x80x2), 5.09 (2H,bs,NH2), 4.22-4.02 (3H,m,NHala+H5xe2x80x2), 3.99-3.87 (1H,m,CHala), 3.59 (3H,t,OCH3), 3.05 (1H,d,H4xe2x80x2), 2.92 (1H,bs,CHcPr), 2.73-2.62 (1H,m,xe2x80x98oxe2x80x99H6xe2x80x2), 1.62-1.53 (1H,m,xe2x80x98oxe2x80x99H6xe2x80x2), 1.30-1.25 (3H,t,CH3ala), 0.78-0.71 (2H,q,2H of CH2cPr), 0.54-0.49 (2H,t,2H of CH2cPr).
13C NMR: xcex4174.6 LCO), 160.3 (C2), 156.6 (C4), 151.3 (C6), 151.1 (xe2x80x98ipsoxe2x80x99-Ph), 136.8 (C8), 135.9 (C2xe2x80x2), 131.5 (C3xe2x80x2), 130.0 (xe2x80x98mxe2x80x99-Ph), 125.2 (xe2x80x98pxe2x80x99-Ph), 120.5 (xe2x80x98oxe2x80x99-Ph), 115.0 (C5), 69.2 (C5xe2x80x2), 59.2 (C1xe2x80x2), 52.8 (OCH3), 50.5 (CHala), 46.0 (C4xe2x80x2), 34.9 (C6xe2x80x2), 24.2 (CHcPr), 21.2 (CH3ala), 7.7 (CH2cPr).
MS ES+: m/z 527.86 (100%) (M)+, 546.84 (M+K)+.
MS FAB: For C24H31O5N7P, requires 528.212431, found 528.213848.
HPLC: tR 30.33 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
IR: 3328.6 (N-Hstr.), 2922.1, 2862.9 (C-Hstr.), 1734.4 (C-Ostr.), 1590.9 (aromatic C-Cstr.), 1462.9 (C-Hdef), 1376.8 (-CH3syM.def.), 1207.1 (P-O-aryl), 1154.0 (C-Ostr.), 1027.7 (P-O-alkyl), 933.4 (olefiric C-Hdef.), 721.8 (monosub.aromatic C-Hdef.).
Phenyl-(methoxy-D-alaninyl)-phosphorochloridate.
C10H13O4N1Cl1P1, MW=277.65.
This was synthesised according to Standard Procedure 3, using D-alanine methyl ester hydrochloride (1.0 g, 7.17 mmol), PhOP(O)Cl2 (1.51 g, 1.07 ml, 7.17 mmol) and NEt3 (1.45 g, 2.0 ml, 14.0 mmol) to yield 1.66 g (83.4%) of crude product that was stored in anhydrous THF (10 ml), to give a 0.60 mmol/ml solution that was used without further purification.
31P NMR: xcex49.38, 9.18 (1:1).
1H NMR: xcex47.39-7.30 (2H,t,xe2x80x98oxe2x80x99-Ph), 7.29-7.09 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 4.85-4.80 (1H,d,NHala), 4.19-4.11 (1H,m,CHala), 3.75 (3H,d,OCH3), 1.52-1.49 (3H,dd,CH3ala).
13C NMR: xcex4173.6 (CO), 150.1 (xe2x80x98ipsoxe2x80x99-Ph), 130.3 (xe2x80x98oxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 120.9 (xe2x80x98mxe2x80x99-Ph), 53.2 (OCH3), 50.9 (CHala), 21.0 (CH3ala).
(1 S,4R)-4-(2-amino-6-cyclopropyiamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl-(methoxy-D-alaninyl)]-phosphate. Cf1583.
C24H30O5N7P1, MW=527.53.
This was synthesised according to Standard Procedure 4, using (1S,4R)1(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (400 mg, 1.4 mmol), tBuMgCl (1.0M solution in THF) (2.1 ml, 2.1 mmol), and phenyl-(methoxy-D-alaninyl)-phosphorochloridate (0.6M solution in THF) (7.0 ml, 4.19 mmol) in THF (25 ml) stirring at room temperature for 36 hrs. The crude product was purified by eluting with 3% MeOH in CHCl3 and then 2.5% MeOH in CHCl3 to give the product as a white foam (318.6 mg, 43.2%).
31p NR: 3.93, 3.70.
1H NMR: xcex47.56+7.51 (1H,d,H8), 7.37-7.32 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.29 (1H,d,xe2x80x98pxe2x80x99-Ph), 7.25-7.15 (2H,m,xe2x80x98mxe2x80x99-Ph), 6.10 (1H,t,J=5.28 Hz,H2xe2x80x2), 6.03 (1H,bs,NHcPr), 5.94-5.89 (1H,m,H3xe2x80x2), 5.54 (1H,bs,H1xe2x80x2), 5.01 (2H,bs,NH2), 4.26-3.83 (4H,m,CHala,NHala+H5xe2x80x2), 3.72 (3H,d,OCH3), 3.18 (1H,s,CHcPr), 3.02 (1H,bs,H4xe2x80x2), 2.86-2.75 (1H,m,1 of H6xe2x80x2), 1.78-1.64 (1H,m,1 of H6xe2x80x2), 1.39-1.36 (3H,dd,CH3ala), 0.90-0.83 (2H,q,J=6.13 Hz,2H of CH2cPr), 0.63 (2H,bs, 2H of CH2cPr).
13C NMR: xcex4174.5 (CO), 160.3 (C2), 156.6 (C4) 151.2 (C6), 151.0 (xe2x80x98ipsoxe2x80x99-Ph), 13608 (C2xe2x80x2), 136.1 (C8), 131.5 (C3xe2x80x2), 130.0 (xe2x80x98mxe2x80x99-Ph), 125.3 (xe2x80x98pxe2x80x99-Ph), 120.5 (xe2x80x98oxe2x80x99-Ph), 115.2 (C5), 69.3 (C5xe2x80x2), 59.3 (Clxe2x80x2), 52.9 (CHala), 50.5 (OCH3), 46.0 (C4xe2x80x2), 34.9 (C6xe2x80x2), 24.1 (CHcPr), 21.4 (CH3ala), 7.8 (CH2cPr).
MS ES+: m/z 527.86 (100%) (M)+, 546.84 (M+K)+.
MS FAB: For C24H31O5N7P requires 528.212431, found 528.211505.
HPLC: tR 29.807 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
IR: 3333.6 (N-Hstr.), 2923.4, 2853.4 (C-Hstr.), 1734.1 (Cxe2x95x90Ostr.), 1591.1 (aromatic C-Cstr.), 1458.3 (C-Hdef.), 1376.7 (-CH3syM.def.), 1208.3 (P-O-aryl), 1153.3 (C-Ostr.), 1026.9 (P-O-alkyl), 931.9 (olefinic C-Hdef), 721.6 (monosub.aromatic C-Hdef).
Phenyl-(methoxy-L-phenylalaninyl)-phosphorochloridate.
C16H17O4N1C1, P1, MW=353.74.
This was synthesised according to Standard Procedure 3, using L-phenylalanine methyl ester (1.0 g, 4.64 mmol), PhOP(O)Cl2 (0.98 g, 0.70 ml, 4.64 mmol) and NEt3 (0.94 g, 1.30 ml, 9.28 mmol) to yield 1.45 g (88.4%) of crude product as an oil that was stored in anhydrous THF (10 ml), to give a 0.41 mmol/ml solution that was used without farther purification. 31p NMR: 6 9.37, 9.23 (1:1).
1H NMR: xcex47.60-7.16 (10H,m,2xPh), 4.704.49 (1H,m,CHala), 4.38-4.16 (1H,m,NHala), 3.89 (3H,d,OCH3), 3.23 (2H,m,CHPh).
(1,S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentee-1-methain O-[phenyl-(methoxy-L-phenylalaninyl)]-phosphate. Cf 1585.
C31H34O5N7P, MW=603.6.
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (300 mg, 1.05 mmol), tBuMgCl (1.0M solution in THF) (1.57 ml, 1.57 mmol) and phenyl-(methoxy-L-phenylalaninyl)-phosphorochloridate (0.41M solution in THF) (7.66 ml, 3.14 mmol) in THF (20 ml) stirring at room temperature for 48 hrs. The crude product was purified by eluting with 3% MeOH in CHCl3 and then 2.5% MeOH in CHCl3 to give the product as a white foam (272.9 mg, 43.15%).
31p NMR: xcex43.91, 3.80.
1H NMR: xcex47.47+7.43 (1H,d,H8), 7.31-7.06 (IOH,m,2xPh), 6.25 (1H,d,NHcPr), 6.00-5.95 (1H,q,H2xe2x80x2), 5.87-5.81 (1H,t,H3xe2x80x2), 5.49 (1H,s,H1xe2x80x2), 5.19 (2H,bs,NH2), 4.31-3.92 (4H,m,CHala,NHala+H5xe2x80x2), 3.64 (3H,d,OCH3), 3.02-2.89 (4H,m,CH2Ph,CHcPr+H4xe2x80x2), 2.78-2.63 (1H,m,1 of H6xe2x80x2), 1.63-1.49 (1H,m,l of H6xe2x80x2), 0.86-0.80 (2H,q,J=6.24 Hz,2H of CH2cPr), 0.60 (2H,d,2H of CH2cPr).
13C NMR: xcex474.3 (CO), 161.5 (C2), 157.7 (C4), 152.4 (C6) 152.1 (xe2x80x98ipsoxe2x80x99-OPh), 137.7 (xe2x80x98ipsoxe2x80x99-Bn), 137.1 (C2xe2x80x2), 136.9 (C8), 132.4 (C3xe2x80x2), 130.9 (xe2x80x98oxe2x80x99+xe2x80x98mxe2x80x99-Bn), 129.9 (xe2x80x98mxe2x80x99-mOPh), 128.4 (xe2x80x98pxe2x80x99-Bn), 126.2 (xe2x80x98pxe2x80x99-OPh), 121.5 (xe2x80x98oxe2x80x99-OPh), 116.1 (C5), 70.1 (C5xe2x80x2), 60.1 (Clxe2x80x2), 57.2 (CHala), 53.6 (OCH3), 46.9 (C6xe2x80x2), 41.7 (C4xe2x80x2), 35.9 (CH2Ph), 25.1 (CHcPr), 8.7 (CH2cPr).
MS ES+: m/z 603.8 (100%, M+), 604.8 (35%, M+H+), 625.7 (15%, M+Na+).
MS FAB: For C31H34O5N7P requires 604.243731, found 604.242585.
HPLC: tR 34.707, 35.020 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
IR: 3331.7 (N-Hstr.), 3007.2, 2952.2 (C-Hstr.), 1741.1 (Cxe2x95x90Ostr.), 1595.6, 1487.7 (aromatic C-Cstr.), 1455.0 (C-Hdef), 1393.9 (xe2x80x94CH3sym.def.), 1252.5 (Pxe2x95x90O), 1214.3 P-0-my1), 1125.3 (C-Ostr.), 1025.6 (P-O-alkyl), 935.8 (olefinic C-Hdef.), 754.8 (monosub.aromatic C-Hdef.).
Phenyl-(methoxyglycinyl)-phosphorochloridate.
C9H11O4N1C1,P1, MW=263.62.
This was synthesised according to Standard Procedure 3, using glycine methyl ester (1.5 g, 11.9 mmol), PhOP(O)Cl2 (2.52 g, 1.79 ml, 11.9 mmol) and NEt3 (2.42 g, 3.33 ml, 23.9 mmol) to yield 3.07 g (97.15%) of crude product as an oil that was stored in anhydrous THF (15 ml), to give a 0.774 mmol/ml solution that was used without further purification.
31p NMR: xcex410.43.
1H NMR: xcex47.43-7.38 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.31-7.25 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 4.67 (1H,bs,NHala), 3.94 (2H,dd,CH2), 3.83 (3H,s,OCH3).
13C NMR: xcex4170.4 (CO), 150.1 (xe2x80x98ipsoxe2x80x99Ph), 130.2 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 120.8 (xe2x80x98oxe2x80x99-Ph), 53.1 (OCH3), 43.4 (CH2).
(1 S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl-(methoxy-glycinyl)]-phosphate. Cf1588.
C23H28O5N7P1, MW=513.49.
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (300 mg, 1.05 mmol), tBuMgCl (1.0M solution in THF) (1.57 ml, 1.57 mmol) and phenyl-(methoxy-glycinyl)-phosphorochloridate (0.774M solution in THF) (4.06 ml, 3.14 mmol) in THF (20 ml) stirring at room temperature for 96 hrs. The crude product was purified by eluting with 3% MeOH in CHCl3 and then with 2.5% MeOH in CHCl3 to give the product as a white foam (82.6 mg, 15.4%).
31p NMR: xcex44.79, 4.67 (1:1).
1H NMR: xcex47.40+7.36 (1H,dH8), 7.24-7.19 (2H,t,xe2x80x98oxe2x80x99-Ph), 7.15-7.10 (2H,t,xe2x80x98mxe2x80x99-Ph), 7.07-7.02 (1H,t,xe2x80x98pxe2x80x99-Ph), 6.00-5.96 (2H,m,H2xe2x80x2+NHcPr), 5.80-5.76 (1H,m,H3xe2x80x2), 5.45-5.41 (1H,tH1xe2x80x2), 4.99 (2H,bs,NH2), 4.14-4.00 (3H,m,NHala+H5xe2x80x2), 3.62 (3H,s,OCH3), 3.03 (1H,d,H4xe2x80x2), 2.91 (1H,d,CHcPr), 2.73-2.62 (1H,m,1of H6xe2x80x2), 1.62-1.51 (1H,m,1 of H6xe2x80x2), 1.45-1.43 (6H,t,2xCH3), 0.78-0.71 (2H,q,2H of CH2cPr), 0.54-0.49 (2H,t,2H of CH2cPr).
13C NMR: xcex4172.1 (CO), 160.2 (C2), 156.6 (C4), 152.0 (C6), 151.7 (xe2x80x98ipsoxe2x80x99-Ph), 137.7 (C8), 137.1 (C2xe2x80x2), 132.0 (C3xe2x80x2), 130.8 (xe2x80x98mxe2x80x99-Ph), 126.0 (xe2x80x98pxe2x80x99-Ph), 121.2 (xe2x80x98oxe2x80x99-Ph), 115.5 (C5), 69.9 (C5xe2x80x2), 60.0 (Clxe2x80x2), 53.5 (OCH3), 46.7 (C4xe2x80x2), 43.9 (CH2), 35.4 (C6xe2x80x2), 25.0 (CHcPr), 8.5 (CH2cPr).
MS ES+: m/z 513.9 (100%, M+), 514.8 (25%, M+H+), 535.8 (40%, M+Na+).
MS FAB: For C23H29O5N7P requires 514.196781, found 514.195321.
HPLC: tR 28.419 (99.9%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
IR: 3342.0 (N-Hstr.), 1749.8 (Cxe2x95x90Ostr.), 1596.2, 1488.4 (aromatic C-Cstr.), 1451.9 (C-Hdef.), 1394.7 (-CH3sym.def), 1259.6 (Pxe2x95x90O), 1212.1 (P-O-aryl), 1151.6 (C-Ostr.), 1026.8 (P-O-alkyl), 937.8 (olefinic C-Hdef), 760.7 (monosub.aromaiic C-Hdef.).
Methyl-2-amino-2-methylpropanoate hydrochloride
C5H,2O2N1Cl, MW=153.61.
This was synthesised according to Standard Procedure 1, using 2-amino-isobutyric acid (4 g, 0.039 mol) with thionyl chloride (5.66 ml, 0.078 mol) and anhydrous methanol (23.5 ml, 0.58 mol). This gave the product as a white solid (5.805 g, 97.4%)
1H NMR (DMSO): xcex48.85 (3H,s,NH+Cl), 3.72 (3H,s,OMe), 1.48 (6H,s,2xMe).
13C NMR (DMSO): xcex4172.8 (COOMe), 56.6 (OMe), 53.9 (CMe2), 24.1 (2xMe).
MS ES+: m/z 117.71 M+H+, 142.88 M+Na+.
Phenyl-(methyl-2-amino-2-methylpropanoate)-phosphorochloridate.
C11I H15O4N1C1P1, MW=291.67.
This was synthesised according to Standard Procedure 3, using 2-amino-isobutate methyl ester hydrochloride (1.0 g, 6.51 mmol), PhOP(O)Cl2 (1.37 g, 0.97 ml, 6.51 mmol) and NEt3 (1.32 g, 1.18 ml, 13.02 mmol), to yield 1.73 g (91%) of the crude product as an oil. This was stored in anhydrous THF (10 ml) to give a solution of 0.593 mmol/ml, and used without further purification.
31P NMR: xcex46.86.
1H NMR: xcex47.43-7.38 (2H,t,xe2x80x98oxe2x80x99-Ph), 7.32-7.21 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 4.84 (1H,d,NHala), 3.83 (3H,s,OCH3), 1.72 (6H,d,2xCH3).
13C NMR: xcex4175.7 (CO), 150.3 (xe2x80x98ipsoxe2x80x99-Ph), 130.3 (xe2x80x98mxe2x80x99-Ph), 126.3 (xe2x80x98pxe2x80x99-Ph), 121.0 (xe2x80x98oxe2x80x99-Ph), 58.8 (OCH3), 53.6 (C[CH3]2), 27.3+27.0 (2xCH3).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol
O-[phenyl-(methoxy-xcex1,adimethylglycinyl)]-phosphate. Cf 1584.
C25H32O5N7P1, MW=542.23.
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (300 mg, 1.05 mmol), tBuMgCl (1.0M solution in THF) (1.57 ml, 1.57 mmol) and phenyl-(methoxy-dimethylglycinyl)-phosphorochloridate (0.59M solution in THF) (5.3 ml, 3.14 mmol) in THF (20 ml) stirring at room temperature for 96 hrs. The crude product was purified by eluting with 3% MeOH in CHCl3 and then with 2.5% MeOH in CHCl3 to give the product as a white foam (193.7 mg, 34.14%).
31p NMR: xcex42.49. 1H NMR: xcex47.40+7.36 (1H,d,H8), 7.24-7.19 (2H,t,xe2x80x98oxe2x80x99-Ph), 7.15-7.10 (2H,t,xe2x80x98mxe2x80x99-Ph), 7.07-7.02 (1H,t,xe2x80x98pxe2x80x99-Ph), 6.00-5.96 (2H,m,H2xe2x80x2+NHcPr), 5.80-5.76 (1H,m,H3xe2x80x2), 5.45-5.41 (1H,t,H1xe2x80x2), 4.99 (2H,bs,NH2), 4.14-4.00 (3H,m,NHala+H5xe2x80x2), 3.62 (3H,s,OCH3), 3.03 (1H,d,H4xe2x80x2), 2.91 (1H,d,CHcPr), 2.73-2.62 (1H,m,1 of H6xe2x80x2), 1.62-1.51 (1H,m,1of H6xe2x80x2), 1.45-1.43 (6H,t,2xCH3), 0.78-0.71 (2H,q,2H of CH2cPr), 0.54-0.49 (2H,t,2H of CH2cycl.).
MS ES+: m/z 541.9 (100%, M+), 563.8 (30%, M+Na+).
MS FAB: For C25H33O5N7P requires 542.228081, found 542.228428.
HPLC: tR 28.347 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
IR: 3346.0 (N-Hstr.), 2923.0, 2853.5 (C-Hstr.), 1734.0 (Cxe2x95x90Ostr.), 1590.2 (aromatic C-Cstr.), 1458.4 (C-Hdef.), 1376.8 (-CH3syn.def.), 1261.3 (Pxe2x95x90O), 1152.7 (C-Ostr.), 1028.0 (P-O-alkyl), 936.0 (olefinic C-Hdef.), 721.7 (monosub.aromatic C-Hdef.).
L-Aspartic acid dimethyl ester hydrochloride.
C6H12O4N1C1, MW=197.62.
This was synthesised according to Standard Procedure 1, using L-asparagine (2.5 g, 0.019 mol) with thionyl chloride (3.67 ml, 0.042 mol) and anhydrous methanol (12.86 ml, 0.32 mol). This gave L-aspartic acid dimethyl ester hydrochloride in 3.70 g, 99% yield. 1H NMR (MeOH-d4): 5 4.53-4.50 (1H,t,CH), 3.94 (3H,s,OCH3), 3.85 (3H,s,OCH3), 3.18 (2H,d,CH2).
13C NMR (MeOH-d): xcex4170.4, 168.4 (CO), 53.0+52.0 (2xOMe), 49.4 (CH), 33.8 (CH2).
Phenyl-(dimethoxy-L-aspartyl)-phosphorochloridate.
C12H15O6N1C1P1, MW=335.68.
This was synthesised according to Standard Procedure 3, using L-Aspartic acid dimethyl ester (1.0 g, 5.04 mmol), PhOP(O)Cl2 (1,06 g, 0.75 ml, 5.04 mmol) and NEt3 (1.02 g, 1.40 ml, 10.1 mmol) to yield 0.55 g (32.4%) of crude product as an oil that was stored in anhydrous THF (5 ml), to give a 0.33 mmol/ml solution that was used without further purification.
31P NMR: xcex49.74,9.59 (1:1).
(1S,4R)-4-(2-amino-6-cyclopropylamin9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl-(L-asparic acid dimethyl ester)]-phosphate. Cf1589.
C24H30O5N7P), MW=527.53.
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (250 mg, 0.87 mol),
tBuMgCl (1.0M solution in THF) (0.87 ml, 0.87 mmol) and phenyl-(L-aspartic acid dimethyl ester)-phosphorochloridate (0.50M solution in THF) (5.20 ml, 2.62 mmol) in THF (15 ml) and stirring at room temperature for 48 hrs. The crude product was purified by eluting with 2.5% MeOH in CHCl3 ((2) to give the product as a pale yellow foam (163.5 mg, 32.0%).
31p NMR: xcex44.19, 3.76 (1:1).
1H NMR: 8 7.40 (1H,d,H8), 7.24-7.19 (2H,t,xe2x80x98oxe2x80x99-Ph), 7.12-7.03 (3H,m,xe2x80x98mxe2x80x99+p-Ph), 6.05-5.95 (2H,m,H2xe2x80x2+NHcPr), 5.79 (1H,d,H3xe2x80x2), 5.44 (1H,s,Hxe2x80x2), 5.02 (2H,bs,NH2), 4.384.07 (4H,m,H5xe2x80x2,NHala+CHala), 3.61 (3H,s,OCH3), 3.54 (3H,d,OCH3), 3.05-2.52 (5H,m,CH2aa,H4xe2x80x2,CHcPr,+1of H6xe2x80x2), 1.64-1.52 (1xe2x80x2H,m,1of H6xe2x80x2), 0.77-0.73 (2H,t,J=5.49 Hz,2Hof CH2cPr), 0.51 (2H,bs,2Hof CH2CPr).
13C NMR: xcex4173.3 (CO), 172.4 (CO), 161.5 (C2), 157.7 (C4), 152.3 (C8), 152.1 (xe2x80x98ispoxe2x80x99-Ph), 137.8 (C2xe2x80x2), 137.0 (C6), 132.6 (C3xe2x80x2), 131.1 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 121.6 (xe2x80x98oxe2x80x99-P3), 116.2 (C5), 70.5 (C5xe2x80x2), 60.3 (Clxe2x80x2), 54.3 (OCH3), 53.5 (OCH3), 52.6 (CHala), 47.1 (C4xe2x80x2), 39.7 (CH2ala), 36.0 (C6xe2x80x2), 25.1 (CHcPr), 8.8 (CH2cPr).
MS ES+: m/z 585.8 (100%, M+), 607.7 (30%, M+Na+.
MS FAB: For C26H33O5N7P requires 586.217910, found 586.217510.
HPLC: tR 29.261 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
IR: 3347.5 (N-Hstr.), 2850.7 (C-Hstr.), 1739.9 (Cxe2x95x90Ostr.), 1596.1 (aromatic C-Cstr.), 1461.9 (C-Hdef.), 1376.6 (xe2x80x94CH3sym.def.), 1262.4 (Pxe2x95x90O), 1211.2 (P-O-aryl), 1158.3 (C-Ostr.), 1027.0 (P-O-alkyl), 935.6 (olefinic C-Hdef), 761.5, 722.0 (monosub.aromatic C-Hdef.).
3-cyclohexyl-L-alanine methyl ester hydrochloride salt
C9H19N1o2Cl1, MW=221.75
This was synthesised according to Standard Procedure 1, using 3-cyclohexyl-L-alanine (3.0 g, 17.5 mmol), methanol (30 ml), and thionyl chloride (2.56 ml, 35 mmol). The product was isolated as a white solid (3.23 g, 83.9%).
1H NMR (MeOH-d4): xcex44.124.07 (3H,t,CHala), 3.85 (3H,s,OCH3), 1.741.68 (6H,m,CH2+0-CH2), 1.56-1.43 (1H,m,CH), 1.36-1.15 (4H,m,m-CH2), 1.05-0.90 (2H,q,p-CH2)
13C NMR: xcex4170.15 (CO), 52.7 (OCH3), 50.8 (CHala), 38.2 (CH2), 33.6 (CH), 33.0+32.7 (2xCH2-o), 26.3 (p-CH2), 26.0+25.9 (2xCH2-m).
Phenyl-(methoxy-3-cyclohexyl-L-alaninyl)-phosphorochloridate
C16H23N1O4P1Cl1, MW=359.82
This was synthesised according to Standard Procedure 3, using 3-Cyclohexyl-L-alanine methyl ester hydrochloride salt (0.7 g, 316 mmol), PhOP(O)Cl2 (0.47 ml, 3.16 mmol), triethylamine (0.88 ml. 6.31 mmol) in DCM (60 ml). The usual workup yielded the crude product as a yellow oil (1.18 g, 100%), which was stored in THF (7 ml) to give a 0.45M solution.
31p NMR. xcex49.79, 9.49 (1:1).
1H NMR: xcex47.49-7.43 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.37-7.19 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 4.464.35 (1H,q,NHala), 4.32-4.20 (1H,m,CHala), 3.88-3.85 (3H,dd,OCH3), 1.94-1.90 (1H,d,CHcHx), 1.76-1.60
(1 S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-Phenyl-(methoxy-3-cyclohexane-L-alaninyl)-phosphate. Cf1709.
C30H40N7O5P1, MW=609.66
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (150 mg, 0.52 mmol), tBuMgCl (1.05 ml, 1.05 mmol, of a 1.0M solution in THF), in THF (4 ml) and phenyl-(methoxy-3-cyclohexane-L-alaninyl)-phosphorochloridate (3.5 ml, 1.57 mmol, of a 0.45M solution in THF), at room temperature for 24 hrs. After 24 hrs, additional phenyl-(methoxy-3-cyclohexane-L-alaninyl)-phosphorochloridate (2.5 ml, 1.12 mmol, of a 0.45M solution in THF) was added and the reaction stirred for another 24 hrs. The crude product was purified by eluting with 3% MeOH in CHCl3, and then 2.5% MeOH in CHCl3 to give the pure product as a pale yellow foamy solid (79.6 mg, 24.9%).
31P NMR: xcex44.14, 3.98 (1:1).
1H NMR: xcex47.50 (1H,d,H8), 7.34-7.13 (5H,t,OPh), 6.20 (1H,s,NHcPr), 6.08 (1H,t,H2xe2x80x2), 5.89 (1H,q,H3xe2x80x2), 5.53 (1H,bs,H1xe2x80x2), 5.16 (2H,bs,NH2), 4.24-3.84 (4H,m,H5xe2x80x2,NHala+CHala), 3.66 (3H,s,OCH3), 3.34 (1H,bs,), 3.11 (1H,d, ), 3.03 (1H,bs, ), 2.84-2.72 (1H,m,1of H6xe2x80x2), 1.98-1.36 (8H,m, ), 1.11 (3H,bs, ), 0.89-0.83 (4H,m,2Hof cPr+4CH2-xe2x80x98pxe2x80x99), 0.63 (2H,d,2Hof cPr).
13C NMR: xcex4174.8CO 160.2 (C2), 156.5 (C4), 151.3 (C6), 151.2 (xe2x80x98ipsoxe2x80x99-Ph), 136.8 (C2xe2x80x2), 135.9 (C8), 131.5 (C3xe2x80x2), 130.0 (xe2x80x98mxe2x80x99-Ph), 125.2 (xe2x80x98pxe2x80x99-Ph), 120.5 (xe2x80x98oxe2x80x99-Ph), 115.1 (C5), 69.4 (C5xe2x80x2), 59.3 (Clxe2x80x2), 52.7 (CHala), 46.1 (C4xe2x80x2), 42.5 (CH2), 34.9 (C6xe2x80x2), 33.8 (CHcHx), 32.7 (CH2-xe2x80x98oxe2x80x99), 26.7 (CH2-xe2x80x98mxe2x80x99), 26.4 (CH2-pxe2x80x2), 24.2 (CHcPr), 7.8 (CH2cPr).
MS ES+: m/z 610.3 (40%, M+), 632.3 (100%, M+Na+), 633.3 (25%, M+H+Na+).
MS FAB: For C30H40O5N7NaP requires 632.2726, found 632.2727.
HPLC: tR 42.154 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-(L-alaninyl)-phosphate diammonium salt Cf1540.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl-(methoxy-L-alaninyl)]-phosphate (125 mg, 0.24 mmol) was stirred in H2O: NEt3 (10 ml, 1:1 v/v), at 25-35xc2x0 C. for 5 hrs. The reaction mixture was extracted with DCM (8xc3x9720 ml), and the aqueous layer reduced to dryness. The resulting solid was solubilised in isopropanol and purified by flash column chromatography, gradient eluting with i-PrOH:H2O: NH3 (11:1:1 to 9:1:2). The appropriate fractions were reduced to dryness and freeze dried to give the pure product as a white foamy solid (106 mg, 95%).
31P NMR (D2O): xcex48.62 (s).
1H N D2O):-xcex47.79 (1H,s,H8), 6.08 (1H,d,H12xe2x80x2), 5.77 (1H,dH3xe2x80x2), 5.35 (1H,t,HIxe2x80x2), 3.71-3.58 (2H,m,H5xe2x80x2), 3.41-3.32 (1H,m,CHa.a), 3.02-2.94 (1H,m,NHCH), 2.70-2.59 (2H,m,H4xe2x80x2+1 of CH2), 1.57-1.49 (1H,dt,l of CH2), 1.10 (3H,d,CH3), 0.83-0.76 (2H,q,1 of CH2cyclo.), 0.61-0.56 (2H,q, 1 of CH2cyclo.).
MS ES+: m/z 437.9 (100%, M+).
MS FAB: calculated m/z 438.165481, found m/z 438.163790.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-(D-alaninyl)-phosphate diammonium salt.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl-(methoxy-D-alaninyl)]-phosphate (100 mg, 0.19 mmol) was stirred in H2O: NEt3 (8 ml, 1:1 v/v), for l 6 hrs. The reaction mixture was extracted with DCM (5xc3x9720 ml), and the aqueous layer reduced to dryness. The resulting solid was solubilised in isopropanol and purified by flash column chromatography, gradient eluting with i-PrOH:H2O:NH3 (11:1:1 to 9:1:2). The appropriate fractions were reduced to dryness and freeze dried to give the pure product as a white foamy solid (88%).
31p NMR (MeOH):-xcex47.81 (s).
1H NMR:-xcex47.74 (1H,s,H8), 6.12 (1H,d,J=5.53 Hz,H2xe2x80x2), 5.78 (1H,t,H3xe2x80x2), 5.44 (1H,d,J=6.21 Hz,H1xe2x80x2), 3.74 (2H,t,J=5.42 Hz,H5xe2x80x2), 3.70-3.60 (1H,m,CHala), 3.01 (1H,bs,H4xe2x80x2), 2.84 (1H,d,J=3.28 Hz,CHcPr), 2.73-2.63 (1H,dt,J=8.66 Hz+5.17 Hz,1of H6xe2x80x2), 1.67-1.58 (1H,m,l of CH2), 1.21 (3H,d,J=7.01 Hz,CH3ala), 0.79-0.73 (2H,q,J=6.68 Hz,2H of CH2cPr), 0.53 (2H,t,2H of CH2cPr).
13C NMR: xcex4179.8 (CO), 161.2 (C2), 157.1 (C4), 151.1 (C6), 139.5 (C2xe2x80x2), 137.8 (C8), 130.7 (C3xe2x80x2), 114.6 (CS), 68.0 (C5xe2x80x2), 60.5 (Clxe2x80x2), 51.9 (CHala), 47.6 (C4xe2x80x2), 35.9 (C6xe2x80x2), 24.4 (CHcPr), 21.7 (CH3ala), 7.6 (CH2cPr).
MS ES+: m/z 437.9 (100%, M+).
MS FAB: calculated m/z 438.165481, found m/z 438.167842.
Phenyl-(ethoxy-L-alaninyl)-phosphorochloridate.
C11H15O4N1Cl1P1, MW=291.67.
This was synthesised according to Standard Procedure 3, using L-Alanine ethyl ester hydrochloride (1.0 g, 6.51 mmol), PhOP(O)Cl2 (1.37 g, 0.97 ml, 6.51 mmol) and NEt3 (1.32 g, 1.81 ml, 13.0 mmol) to yield 1.85 g (97.4%) of crude product as an oil that was stored in anhydrous THF (10 ml), to give a 0.63 mmol/ml solution that was used without further purification.
31P NMR: xcex49.41, 9.16 (1:1).
1H NMR: xcex47.42-7.35 (2H,dd, xe2x80x98oxe2x80x99-Ph), 7.31-7.25 (3H,m, xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 4.71 (1H,d,NHala), 4.314.13 (3H,m,OCH2+CHala), 1.55-1.52 (3H,dd,OCH CH3), 1.33-1.30 (3H,dd,CH3ala).
13C NMR: xcex4173.1 (CO), 150.2 (xe2x80x98ipsoxe2x80x99-Ph), 130.3 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 120.9 (xe2x80x98oxe2x80x99-Ph), 62.3 (OCH2), 51.0 (CHala), 20.9 (CH2CH3), 14.5 (CH3ala).
(1S,4R)-4-(2-amino-6cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methaanol O-[phenyl-(ethoxy-L-alaninyl)]-phosphate. Cf1587.
C24H30 O5N7P1, MW=527.53.
This was synthesised according to Standard Procedure 4, using (1S,4R)-42-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (300 mg, 1.4 mmol), tBuMgCl (1.0M solution in THF) (1.57 ml, 1.57 mmol), and phenyl-(ethoxy-L-alaninyl) phosphorochloridate (0.49M solution in THF) (6.45 ml, 3.14mmol) in anhydrous THF (20 ml), and stirring at room temperature for 24 hrs. The crude product was purified by column chromatography eluting with 2.5% MeOH in CHCl3 to give the product as a pale yellow foam (290 mg, 51.1%).
31p NMR: xcex44.04, 3.96 (1:1).
1H N: xcex47.39 (1H,d,J=7.56 Hz,H8), 7.23-7.18 (2H,t,J=7.90 Hz,xe2x80x98oxe2x80x99-Ph), 7.12-7.10 (2H,t,xe2x80x98mxe2x80x99-Ph), 7.06-7.01 (1H,t,J=7.13 Hz,xe2x80x98pxe2x80x99-Ph), 6.18 (1H,bs,NHcPr), 5.97-5.95 (1H,tH2xe2x80x2), 5.79-5.75 (1H,t,J=5.55 Hz,H3xe2x80x2), 5.43 (1H,s,H1xe2x80x2), 5.13 (2H,bs,NH2), 4.30-4.14 (1H,m,NHala), 4.064.00 (4H,m,H5xe2x80x2+OCH2), 3.96-3.84 (1H,m,CHala), 3.03 (1H,d,J=5.74 Hz,H4xe2x80x2), 2.92 (1H,bs,CHcPr), 2.71-2.61 (1H,m,1of H6xe2x80x2), 1.60-1.51 (1H,m, 1of H6xe2x80x2), 1.29-1.24 (3H,tJ=6.64 Hz,CH3ala), 1.18-1.11 (3H,m,CH2CH3), 0.75-0.71 (2H,q,J=6.76 Hz,2H of CH2cPr), 0.50 (2H,bs,2H of CH2cPr).
13C NMR: xcex4173.35 (CO), 159.8 (C2), 156.0 (C4), 150.6 (C6) 150.4(xe2x80x98ipsoxe2x80x99-Ph), 136.1 (C2xe2x80x2), 135.1 (C8), 130.8 (C3xe2x80x2), 129.3 (xe2x80x98mxe2x80x99-Ph), 124.5 (xe2x80x98pxe2x80x99-Ph), 119.8 (xe2x80x98oxe2x80x99-Ph), 114.4 (C5), 68.6 (C5xe2x80x2), 61.2 (OCH2), 58.5 (Clxe2x80x2), 50.0 (CHala), 45.3 (C4xe2x80x2), 34.3 (C6xe2x80x2), 23.4 (CcPr), 20.6 (CH3ala), 13.8 (CH2CH3), 7.0 (CH2cPr).
MS ES+: m/z 541.9 (100%, M+), 546.84 (28%, M+H+), 563.8 (25%, M+Na+).
MS FAB: For C25H33O5N7P, requires 542.228081, found 542.228131.
HPLC: tR 31.76, 32.03 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
IR: 3334.1 (N-Hstr.), 1734.5 (Cxe2x95x90Ostr.), 1595.9, 1488.0 (aromatic C-Cstr.), 1450.3 (C-Hdef.), 1394.2 (-CH3sym.def.), 1252.8 (Pxe2x95x90O), 1210.4 (P-O-aryl), 1153.3 (C-Ostr.), 1026.0 (P-O-alkyl), 934.8 (olefinic C-Hdef.), 759.0 (monosub.aromatic C-Hdef.).
Phenyl-(benzoxy-L-alaninyl)-phosphorochlioridate.
C16H17O4N1Cl1P1, MW=353.74.
This was synthesised according to Standard Procedure 3,using L-alanine benzyl ester hydrochloride (1.0 g, 4.64 mmol), PhOP(O)Cl2 (0.98 g, 0.69 ml, 4.64 mmol) and NEt3 (0.94 g, 1.29 ml, 9.27 mmol) to yield 1.61 g (98.2%) of crude product that was stored in anhydrous THF (10ml), to give a 0.46 mmol/ml solution that was used without further purification.
31p NMR: xcex49.41, 9.23 (1:1).
1H NMR: xcex47.41-7.21 (10H,m,2xPh), 5.24 (2H,d,CH2Ph), 4.95-4.88 (1H,t,NHala), 4.36-4.15 (1H,m,CHala), 1.56 (3H,t,CH3ala).
13C NMR: xcex4172.9 (CO), 150.2 (xe2x80x98ipsoxe2x80x99-OPh), 135.5 (xe2x80x98ipsoxe2x80x99-CH2Ph), 130.3 (xe2x80x98mxe2x80x99-OPh), 129.0 (xe2x80x98oxe2x80x99-CH2Ph), 128.7 (xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-CH2Ph), 126.4 (xe2x80x98pxe2x80x99-OPh), 121.0 (xe2x80x98oxe2x80x99-OPh), 68.0 (OCH2), 51.1 (CHala), 20.8 (CH3ala).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl-(enzoxy-L-alaninyl)]-phosphate. Cf1582.
C30H35O5N7P1, MW=603.6.
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (400 mg, 1.44 mmol), tBuMgCl (1.0M solution in THF) (2.1 ml, 2.1 mmol), and phenyl-(benzoxy-L-alaninyl)-phosphorochloridate (0.46M solution in THF) (9.2 ml, 4.19 mmol) in anhydrous THF (20 m1), and stirring at room temperature for 64 hrs. The crude product was purified by column chromatography eluting with 3% MeOH in CHCl3, and then 2.5% MeOH in CHCl3 to give the product as a white foam (82.2 mg, 9.75%).
A second synthesis was undertaken with (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (200 mg, 0.7 mmol), tBuMgCl (2.43 ml of a 1.0M soln in THF, 2.43 mmol), and phenyl-(benzoxy-L-alaninyl)-phosphorochloridate (2.2 ml of a 0.46M soln in THF, 2.1 mmol) in THF (2.5 ml). Purification by column chromatography eluting with 3% MeOH in CHCl3 gave the pure product as a white foamy solid (90 mg, 21.3%).
31P NMR: 8 3.82, 3.72 (1:1). 1H NMR: xcex47.51 (1H,d,H8), 7.37-7.15 (IOH,m,OPh+CH2Eh), 6.10-6.04 (1H,m,H2xe2x80x2), 5.96 *(H,bs,NHcPr), 5.89 (1H,dd,J=5.36 HzH3xe2x80x2), 5.54 (1H,t,H1xe2x80x2), 5.16 (2H,bs,NH2), 4.96 (2H,bs,CH Ph), 4.234.05 (3H,m,NHala+H5xe2x80x2), 3.89-3.70 (1H,dt,CHala), 3.16-3.12 (1H,t,H4xe2x80x2), 3.03 (1H,bs,CHcPr), 2.85-2.71 (1H,m,1of H6xe2x80x2), 1.74-1.64 (1H,m,1of H6xe2x80x2), 1.44-1.39 (3H,t,J=7.84 Hz,CH3ala), 0.88 (2H,q,J=6.75 Hz,2H of CH2cPr), 0.64 (2H,m,2H of CH2cPr).
13C NMR: xcex4173.3 (CO), 159.7 (C2), 156.0 (C4), 150.9 (C6), 150.7 (xe2x80x98ipsoxe2x80x99-OPh), 136.4 (C2xe2x80x2), 135.7 (xe2x80x98ipsoxe2x80x99-Bn), 135.2 (CS), 131.0 (C3xe2x80x2), 129.6 (xe2x80x98oxe2x80x99-Bn), 128.6 (xe2x80x98mxe2x80x99-Bn), 128.5 (xe2x80x98pxe2x80x99-Bn), 128.2 (xe2x80x98mxe2x80x99-OPh), 124.9 (xe2x80x98pxe2x80x99-OPh), 120.1 (xe2x80x98oxe2x80x99-OPh), 114.8 (C5), 68.8 (C5xe2x80x2), 67.2 CH2Ph), 58.9 (Clxe2x80x2), 50.3 (CHala), 45.6 (C4xe2x80x2), 34.4 (C6xe2x80x2), 23.7 (CHcPr), 21.0 (CH3ala), 7.4 (CH2cPr).
MS ED+: m/z 603.8 (100%, M+), 604.8 (30%, M+H+), 625.7 (20%, M+Na+).
MS FAB: For C30H35O5N7P requires 604.243731, found 604.241775.
HPLC: tR 33.39 (99.7%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
IR: 3355.9 (N-Hstr.), 2923.3, 2853.7 (C-Hstr.), 1734.1 (Cxe2x95x90Ostr.), 1595.6 (aromatic C-Cstr.), 1458.4 (C-Hdef.), 1376.5 (-CH3sym.def.), 1154.4 (C-Ostr.), 1028.2 (P-O-alkyl), 935.8 (olefinic C-Hdef.), 721.7 (monosub.aromatic C-Hdef).
L-Alanine n-propyl ester hydrochloride salt.
C6H14N1O2Cl1, MW=167.634
This was synthesised according to Standard Procedure 1, using anhydrous propan-1-ol (42.0 ml, 0.56 mol), thionyl chloride (8.2 ml, 0.112 mol) and L-alanine (5.0 g, 0.056 mol). The product was isolated as a white solid (8.88 g, 94.3%).
1H NMR (MeOH-d4): xcex44.344.26 (2H,m,OCH2), 4.244.17 (1H,q,CHala), 1.88-1.78 (2H,m,CH2), 1.65 (3H,d,J=7.24 Hz,CH3ala), 1.10-1.05 (3H,t,CH2CH3). 13C NMR: xcex4170.1 (CO), 68.0 (OCH2), 48.9 (CHala), 21.9 (CH2), 15.3 (CH3ala), 9.5 (CH2CH3)
Phenyl-(n-propoxy-L-alaninyl)-phosphorochloridate
C12H17N1O4P1Cl1, MW=305.79
This was synthesised according to Standard Procedure 3, using L-Alanine n-propyl ester hydrochloride salt (0.5 g, 2.98 mmol), PhOP(O)Cl2 (0.45 ml, 2.98 mmol), triethylamine (0.83 ml. 5.97 mmol) in DCM (70 ml). The usual workup yielded the crude product as a yellow oil (0.84 g, 92.1%), which was stored in THF (5 ml) to give a 0.55M solution.
31P NMR: xcex49.41, 9.17 (1:1).
13C NMR: xcex4173.1 (CO), 150.1 (xe2x80x98ipsoxe2x80x99-Ph), 130.0 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 121.0 (xe2x80x98oxe2x80x99-Ph), 67.9 (OCH2), 51.0 (CHala), 22.3 (CH2CH3), 21.0 (CH3ala), 10.7 (CH,CH3).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-Sl)-2-cyclopentene-1-methanol O-Phenyl-(n-propoxy-L-alaninyl)-phosphate. Cf1646.
C26H34N7O5P, MW=555.57
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (100 mg, 0.35 mmol), tBuMgCl (0.7 ml, 0.7 mmol, of a 1.0M solution in THF), in THF (3 ml) and phenyl-(n-propyl-L-alaninyl)-phosphorochloridate (1.9 ml, 1.05 mmol, of a 0.55M solution in THF), at room temperature for 24 hrs. The crude product was purified by eluting with 3% MeOH in CHCl3 to give the pure product as a pale yellow foamy solid (123 mg, 63.4%).
31P NMR: xcex44.06, 3.98 (1:1).
1H NMR: xcex47.40 (1H,d,J=7.99 HzH8), 7.23-7.18 (2H,dd,xe2x80x98oxe2x80x99-Ph), 7.12-7.02 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 6.16 (1H,bs,H3xe2x80x2), 5.96 (1H,tH2xe2x80x2), 5.78 (1H,d,J-5.83 Hz,NHcycl), 5.44 (1H,bs,H1xe2x80x2), 5.15 (2H,bs,NH2), 4.33-4.18 (1H,m,CHala), 4.15-4.04 (2H,m,OCH2), 4.01-3.88 (2H,m,H5xe2x80x2), 3.65 (1H,bs,NHala), 3.03 (1H,d,H4xe2x80x2), 2.92 (1H,bs,CHcycl), 2.72-2.62 (1H,m,1of H6xe2x80x2), 1.60-1.47 (3H,m,1of H6xe2x80x2+CH2CH3), 1.30-1.26 (3H,t,CH3ala), 0.84-0.80 (3H,m,CHCH:), 0.73 (2H,d,J=6.8 Hz,1of CH2cycl), 0.51 (2H,bs,1 of CH2cycl).
13C NMR: xcex4174.ICO 160.4 (C2), 156.6 (C4), 151.1 (C6+xe2x80x98ipsoxe2x80x99-Ph), 136.8 (C2xe2x80x2), 135.9 (C8), 131.5 (C3xe2x80x2), 130.0 (xe2x80x98mxe2x80x99-Ph), 125.2 (xe2x80x98pxe2x80x99-Ph), 120.5 (xe2x80x98oxe2x80x99-Ph), 115.0 (C5), 69.2 (C5xe2x80x2), 67.4 (OCH2), 59.2 (Clxe2x80x2), 50.6 (CHala), 46.0 (C4xe2x80x2), 35.0 (C6xe2x80x2), 24.2 (CHcPr), 22.3 (CH2CH3), 21.5 (CH3ala), 10.7 (CH2CH3), 7.7 (CH2cycl).
MS ED+: m/z 555.8 (100%, M+), 557.0 (30%, M+H+).
MS FAB: For C26H35O5N7P requires 556.2437, found 556.2438.
HBLC: tR 34.708 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
L-Alanine n-butyl ester hydrochloride salt.
C7H16N1O2Cl1, MW=181.661
This was synthesised according to Standard Procedure 1, using anhydrous butan-1-ol (51.4 ml, 0.56 mol), thionyl chloride (8.2 ml, 0.l 12 mol) and L-alanine (5.0 g, 0.056 mol). The product was isolated as a white solid (8.86 g, 86.9%).
1H NMR (MeOH-d4): xcex44.29-4.17 (2H,m,OCH2), 4.13-4.06 (1H,q,CHala), 1.71-1.62 (2H,m,OCH2CH2), 1.53 (3H,d,J=7.25 Hz,CH3ala), 1.47-1.34 (2H,m,CH2CH3), 0.96-0.91 (3H,t,CH CH).
13C NMR: xcex4170.1 (CO), 66.2 (OCH2), 48.9 (CHala), 30.6 (OCH2CH2), 19.0 (CH2CH3), 15.3 (CH3ala), 13.0 (CH2CH2).
Phenyl-(n-butoxy-L-alaninyl)-phosphorochloridate
C13H19N1O4P1Cl1, MW=337.82
This was synthesised according to Standard Procedure 3, using L-Alanine n-butyl ester 15 hydrochloride salt (0.5 g, 2.75 mmol), PhOP(O)Cl2 (0.41 ml, 2.75 mmol), triethylamine (0.77 m1.5.5 mmol) in DCM (80 ml). The usual workup yielded the crude product as a yellow oil (0.84 g, 94.5%), which was stored in THF (5 ml) to give a 0.525M solution.
31P NMR: xcex49.39, 9.10 (1:1).
1H NMR: xcex47.43-7.15 (5H,m,Ph), 4.68-4.59 (1H,q,CHala), 4.27-4.05 (3H,m,OCH2+NHala), 1.73-1.59 (2H,m,OCH2CH2), 1.56-1.53 (2H,dd,CH2CH3), 1.46-1.37 (3H,m,CH3ala), 1.00-0.92 (3H,m,CH2CH3).
13C NMR: xcex4173.2 (CO), 150.1 (xe2x80x98ipsoxe2x80x99-Ph), 130.3 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 121.0 (xe2x80x98oxe2x80x99-Ph), 66.2 (OCH2), 51.0 (CHala), 30.9 (OCH2CH7), 21.0 (CH3ala), 19.4 (CH2CH3), 14.1 (CH2CH3).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-Phenyl-(n-butoxy-L-alaninyl)-phosphate. Cf1647.
C27H36N7O5P1, MW=569.597
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (100 mg, 0.35 mmol), tBuMgCl (0.7 ml, 0.7 mmol, of a 1.0M solution in THF), in THF (3 ml) and phenyl-(n-butyl-L-alaninyl)-phosphorochloridate (2.0 ml, 1.05 mmol, of a 0.525M solution in THF), at room temperature for 24 hrs. The crude product was purified by eluting with 3% MeOH in CHCl3 to give the pure product as a pale yellow foamy solid (157 mg, 78.9%).
31P NMR: xcex44.01, 3.95 (1:1).
1H NMR: xcex47.40 (1H,d,J=7.32 Hz,H8), 7.23-7.18 (2H,t,xe2x80x98oxe2x80x99-Ph), 7.11 (2H,t,xe2x80x98mxe2x80x99-Ph), 7.04 (1H,t,xe2x80x98pxe2x80x99-Ph), 6.02 (1H,bs,H3xe2x80x2), 5.97 (1H,t,H2xe2x80x2), 5.78 (1H,bs,NHcycl), 5.44 (1H,bs,H1xe2x80x2), 5.06 (2H,bS,NH2), 4.22-3.88 (6H,m,CHala,OCH2,H5xe2x80x2+NHala), 3.05 (1H,d,H4xe2x80x2), 2.93 (1H,bs,CHcycl), 2.72-2.62 (1H,m,1of H6xe2x80x2), 1.61-1.47 (3H,m,1of H6xe2x80x2+OCH7CH ), 1.30-1.26 (5H,t,CH3ala+CH2CH3), 0.85-0.80 (3H,t,CH CH), 0.74 (2H,d,J=6.45 Hz,1of CH2cycl), 0.51 (2H,bs,1of CH2cycl).
13C NMR: xcex4174.1(CO), 160.4 (C2), 156.7 (C4), 151.2 (C6), 151.1 (xe2x80x98ipsoxe2x80x99-Ph), 136.7 (C2xe2x80x2), 135.8 (C8), 131.5 (C3xe2x80x2), 130.0 (xe2x80x98mxe2x80x99-Ph), 125.2 (xe2x80x98pxe2x80x99-Ph), 120.5 (xe2x80x98oxe2x80x99-Ph), 115.0 (C5), 69.3 (C5xe2x80x2), 65.8 (OCH2), 59.2 (Clxe2x80x2), 50.6 (CHala), 46.0 (C4xe2x80x2), 35.0 (C6xe2x80x2), 30.9 (OCH2CH2), 24.1 (CHcPr), 21.5 (CH3ala), 19.4 (CH2CH3), 14.1 (CH2CH3), 7.8 (CH2Cycl).
MS ES+: m/z 569.9 (70%, M+), 570.9 (20%, M+H+), 591.8 (100%, M+Na+), 607.8 (20%, M+K+).
HPLC: tR 388.27 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
L-Alanine i-propyl ester bydrochloride salt.
C6H14N1O2Cl1, MW=167.634
This was synthesised according to Standard Procedure 1, using anhydrous propan-2-ol (43.0 ml, 0.56 mol), thionyl chloride (8.2 ml, 0.1 12 mol) and L-alanine (5.0 g, 0.056 mol). The product was isolated as a semicrystalline solid (8.86 g, 86.9).
1H NMR (MeOH-d4): xcex45.16-5.08 (1H,m,CHala), 4.11-4.04 (1H,q,OCH(Me)2), 1.55 (3H,d,J-7.2 1 Hz,CH3ala), 1.34-1.31 (6H,dd,CH(Me)2). 13C NMR: 6 169.5 (CO), 70.8 (COCH(Me)2), 48.9 (CHala), 20.8 (CH3ala), 15.3 (CH(Me)2).
Phenyl-(i-propoxy-L-alaninyl)-phosphorochloridate
C12H17N1O4P1Cl1, Mw=305.79
This was synthesised according to Standard Procedure 3, using L-Alanine i-propyl ester hydrochloride salt (0.5 g, 2.98 mmol), PhOP(O)Cl2 (0.45 ml, 2.98 mmol), triethylamine (0.83 ml. 5.97 mmol) in DCM (70 ml). The usual workup yielded the crude product as a yellow oil (1.12 g,  greater than 100%), which was stored in THF (5 ml) to give a 0.597M solution.
31P NMR: xcex49.45, 9.17 (1:1).
13C NMR: xcex4172.6 (CO), 150.2 (xe2x80x98ipsoxe2x80x99-Ph), 130.3 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 121.0 (xe2x80x98oxe2x80x99-Ph), 70.1 (OCH), 51.1 (CHala), 22.1 (CH(CH3)2), 20.9 (CH3ala).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-Phenyl-(i-propoxy-L-alaninyl)-phosphate Cf1661.
C26H34N7O5P1, MW=555.57
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (100 mg, 0.35 mmol), tBuMgCl (0.7 ml, 0.7 mmol, of a 1.0M solution in THF), in THF (3 ml) and phenyl-(i-propyl-L-alaninyl)-phosphorochloridate (1.76 ml, 1.05 mmol, of a 0.597M solution in THF), at room temperature for 72 hrs. The crude product was purified by eluting with 3% MeOH in CHCl3 (x2) to give the pure product as a pale yellow foamy solid (106.8 mg, 54.8%).
31P NMR: xcex44.02, 3.98 (1:1).
1H NMR: xcex47.41 (1H,d,J=8.12 Hz,H8), 7.24-7.19 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.13-7.03 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 6.37 (1H,bs,CHcPr), 5.98 (1H,t,H3xe2x80x2), 5.80-5.76 (1H,m,H2xe2x80x2), 5.43 (H,bs,H1xe2x80x2), 5.21 (2H,bs,NH2), 4.944.86 (1H,m,OCH), 4.15-3.98 (2H,m,H5xe2x80x2), 3.92-3.83 (1H,m,CHala), 3.59 (1H,bs,NHala), 3.06-2.98 (1H,m,H4xe2x80x2), 2.93 (1H,bs,CHCPr), 2.74-2.63 (1H,m,1of H6xe2x80x2), 1.62-1.53 (1H,m,1of H6xe2x80x2), 1.34-1.18 (3H,m,CH3ala), 1.15-1.11 (6H,m,CH(CH3)2), 0.79-0.73 (2H,q,2Hof CH2cPr), 0.53 (2H,bs,2Hof CH2cPr).
13C NMR: xcex4173.5(CO) 159.8 (C2), 156.2 (C4), 151.1 (C6), 151.0 (xe2x80x98ipsoxe2x80x99-Ph), 136.9 (C2xe2x80x2), 136.1 (C8), 131.3 (C3xe2x80x2), 130.0 (xe2x80x98mxe2x80x99-Ph), 125.3 (xe2x80x98pxe2x80x99-Ph), 120.5 (xe2x80x98oxe2x80x99-Ph), 115.0 (C5), 69.6 (C5xe2x80x2), 69.2 (OCH), 59.3 (Clxe2x80x2), 50.7 (CHala), 46.0 (C4xe2x80x2), 34.9 (C6xe2x80x2), 24.2 (CHcPr), 22.0 (CH(CH3)2)2), 21.4 (CH3ala), 7.8 (CH2cycl).
xe2x96xa1S ES+: m/z 555.9 (100%, M+), 556.9 (30%, M+H+).
MS MALD/I TOF: For C26H35O5N7P found 555.575.
HPLC: tR 35-85 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
Phenyl-tertbutyloxy-L-alaninyl phosphorochloridate.
C16H17O4N1Cl1P1, MW=353.74.
This was synthesised according to Standard Procedure 3, using L-alanine tert-butyl ester hydrochloride (0.5 g, 2.75 mmol), PhOP(O)Cl2 (0.41 ml, 2.75 mmol) and NEt3 (0.77 ml, 5.5 mmol) to yield 0.77 g (87.5%) of crude product that was stored in anhydrous THF (5 ml), to give a 0.48 mmol/ml solution that was used without further purification.
31P NMR: xcex49.53, 9.20 (1:1).
1H NMR: xcex47.44-7.39 (2H,t,xe2x80x98oxe2x80x99-Ph), 7.32-7.26 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 4.4714.34 (1H,m,NHala), 4.17-4.04 (1H,m,CHala), 1.53 (9H,3s,3xCH3).
13C NMR: xcex4170.7 (CO), 148.7 (xe2x80x98ipsoxe2x80x99-Ph), 128.9 (xe2x80x98oxe2x80x99-Ph), 124.9 (xe2x80x98pxe2x80x99-Ph), 119.5 (xe2x80x98mxe2x80x99-Ph), 81.65 (CMe3), 50.0 (CHala), 26.9 (3xCH3).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-(phenyl-tertbutyloxy-L-alaninyl)-phosphate. Cf1645.
C24H30 O5N7P1, MW=603.6.
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (140 mg, 0.52 mmol), tBuMgCl (1.05 ml, 1.05 mmol of a 1.0M solution in THF), and phenyl-(tertbutyloxy-L-alaninyl)-phosphorochloridate (3.3 ml, 1.57 mmol, of a 0.48M solution in THF), in anhydrous THF (4 ml) stirring at room temperature for 48 hrs. The crude product was purified by eluting with 3% MeOH in CHCl3 to give the pure product as white foamy solid (192.3 mg, 69.0%).
31P NMR: xcex44.15 (s).
1H NMR: xcex47.40 (1H,d,J=8.35 Hz,H8), 7.23-7.18 (2H,t,xe2x80x98mxe2x80x99-Ph), 7.12 (2H,d,xe2x80x98oxe2x80x99-Ph), 7.06-7.02 (1H,t,xe2x80x98pxe2x80x99-Ph), 6.09 (1H,bs,H2xe2x80x2), 5.97 (1H,bs,H3xe2x80x2), 5.77 (1H,d,NHcPr), 5.44 (1H,bs,HIxe2x80x2), 5.10 (2H,bs,NH2), 4.144.05 (3H,m,H5xe2x80x2+NHala), 3.85-3.77 (1H,q,CHala), 3.04 (1H,bs,H4xe2x80x2), 2.93 (1H,bs,CHcPr), 2.72-2.62 (1H,m,1of H6xe2x80x2), 1.58-1.53 (1H,t,1of H6xe2x80x2), 1.34 (9H,d,CMe3), 1.27-1.23 (3H,t,CH3ala), 0.73 (2H,d,2Hof CH2cPr), 0.51 (2H,bs,2Hof CH2cPr).
13C NMR: xcex4173.2 (CO), 160.4 (C2), 156.7 (C4), 151.2 (C6+xe2x80x98ipsoxe2x80x99-Ph), 136.8 (C2xe2x80x2), 135.9 (C8), 131.5 (C3xe2x80x2), 130.0 (xe2x80x98mxe2x80x99-Ph), 125.2 (xe2x80x98pxe2x80x99-Ph), 120.6 (xe2x80x98oxe2x80x99-Ph), 115.2 (C5), 82.3 (C[H3]3), 69.3 (C5xe2x80x2), 59.1 (Clxe2x80x2), 46.0 (C4xe2x80x2), 35.0 (C6xe2x80x2), 28.3 (3xCH3), 24.2 (CHCPr), 1.5 (CH3ala), 7.8 (CH2cPr).
S ES+: m/z 570.0 (100%, M+), 570.9 (32%, M+H+).
MS FAB: For C27H37O5N7P requires 570.2594, found 570.2598.
HPLC: tR 36.158 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
L-Alanine n-pentyl ester hydrochloride salt
C8H16N1O2Cl1, MW=195.69
This was synthesised according to Standard Procedure 1, using pentanol (36.3 ml, 0.337 mol), thionyl chloride (4.92 ml, 67.4 mmol) and L-Alanine (3.0 g, 33.7 mmol). The product was isolated as a white solid pure product (4.86 g, 73.7%).
1H NMR (MeOH-d4): xcex44.324.20 (2H,m,OCH2), 4.16-4.08 (1H,m,CHala), 1.77-1.68 (2H,m,OCH2CH2), 1.56 (3H,d,J=7.22 Hz,CH3ala), 1.42-1.36 (4H,m,CH2CH2CH3), 0.97-0.93 (3H,m,CH2CH3).
13C NMR: xcex4170.1 (CO), 66.5 (OCH2), 48.8 (CHala), 28.2 (OCH2CH2), 28.0 (CH2CH2CH3), 22.3 (CH2CH3), 15.2 (CH3ala), 13.3 (CH2CH3).
Phenyl-(n-pentoxy-L-alaninyl)-phosphorochloridate
C14H21N1O4P1Cl1, MW=333.78
This was synthesised according to Standard Procedure 3, using L-Alanine n-pentyl ester hydrochloride salt (0.5 g, 2.56 mmol), PhOP(O)Cl2 (0.38 ml, 2.56 mmol), triethylamine (0.71 ml. 5.11 mmol) in DCM (60 ml). The usual workup yielded the crude product as a yellow oil (0.79 g, 92.6%), which was stored in THF (5 ml) to give a 0.47M solution.
31P NMR: xcex49.39, 9.12 (1:1).
1H NMR: xcex47.43-7.38 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.32-7.25 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 4.63 (1H,bd,NHala), 4.244.11 (3H,m,OCH2+CHala), 1.73-1.65 (2H,m,OCH2CH,), 1.57-1.53 (3H,dd,CH3ala), 1.42-1.35 (4H,m,2xCH2), 0.97-0.91 (3H,m,CH2CH3).
13C NMR: xcex4173.1 (CO), 150.1 (xe2x80x98ipsoxe2x80x99-Ph), 130.3 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 121.0 (xe2x80x98oxe2x80x99-Ph), 66.5 (OCH2), 51.0 (CHala), 28.6 (CH2-C2), 28.3 (CH2-C3), 22.7 (CH2-C4), 21.0 (CH3ala), 14.1 (CH3-C5).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol 0-Phenyl-(n-pentyloxy-L-alaninyl)-phosphate. Cf1706.
C28H38N7O5P1, MW=583.7
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (10 mg, 0.35 mmol), tBuMgCl (0.7 ml, 0.7 mmol, of a 10M solution in THF), in THF (3 ml) and phenyl-n-pentyl-L-alaninyl)-phosphorochloridate (2.22 ml, 1.05 mmol, of a 0.47M solution in THF), at room temperature for 24 hrs. The crude product was purified by eluting with 2.5-3.0% MeOH in CHCl3 (x2) to give the pure product as a pale yellow foamy solid (143.2 mg, 70.2%).
31P NMR: xcex43.99, 3.95 (1:1).
1H NMR: xcex47.41 (1H,d,J=7.18 Hz,H8), 7.24-7.19 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.12-7.02 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 6.09 (1H,bs,NHcPr), 5.98 (1H,d,H2xe2x80x2), 5.79 (1H,bs,H3xe2x80x2), 5.44 (1H,bs,H1xe2x80x2), 5.09 (2H,bs,NH2), 4.16-3.88 (6H,m,CHala,OCH2,H5xe2x80x2+NHala), 3.05 (1H,bs,H4xe2x80x2), 2.94 (1H,bs,CHcPr), 2.73-2.63 (1H,m,1of H6xe2x80x2), 1.62-1.51 (3H,m,1of H6xe2x80x2+OCH2CH2), 1.31-1.21 (7H,t,CH3ala+2xCH2), 0.81-0.74 (5H,m,CH3+2Hof CH2cPr), 0.52 (2H,bs,2Hof CH2cPr).
13C NMR: xcex4174.1(CO), 160.1 (C2), 156.5 (C4), 151.2 (C6), 151.1 (xe2x80x98ipsoxe2x80x99-Ph), 136.8 (C2xe2x80x2), 136.1 (C8), 131.5 (C3xe2x80x2), 130.0 (xe2x80x98mxe2x80x99-Ph), 125.2 (5xe2x80x98pxe2x80x99-Ph), 120.5 (xe2x80x98oxe2x80x99-Ph), 115.1 (C5), 69.3 (C5xe2x80x2), 66.1 (OCH2), 59.3 (Clxe2x80x2), 50.7 (CHala), 46.0 (C4xe2x80x2), 34.9 (C6xe2x80x2), 28.6 (CH2-C2), 28.3 (CH2-C3), 24.2 (CHcPr), 22.6 (CH2-C4), 21.5 (CH3ala), 14.3 (CH3-C5), 7.8 (CH2cPr).
MS ES+: m/z 584.2 (100%, M+), 585.2 (25%, M+H+).
MS FAB: For C28H39O5N7P requires 584.2750, found 584.2757.
BPLC: tR 40.294 (99.3%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
L-Alanine in-hexyl ester bydrochloride salt
C9H20N1O2Cl1, MW=209.75
This was synthesised according to Standard Procedure 2, using L-Alanine (2.0 g, 22.5mmol), hexan-1-ol (2.82 ml, 22.5 mmol), p-toluene sulfonic acid monohydrate (4.7 g, 24.7 mmol), and toluene (100 ml). L-alanine n-hexyl ester hydrochloride was isolated as a white powdery solid (3.32 g, 70.5%).
1H NMR (MeOH-d4): 5 4.31-4.18 (2H,m,OCH2), 4.17-4.09 (1H,q,CHala), 1.75-1.66 (2H,m,OCH2CH2), 1.57 (3H,d,J=7.20 Hz,CH3ala), 1.45-1.35 (6H,m,[CH2]3CH3), 0.94-0.89 (3H,t,CH2CH3).
13C NMR: xcex4170.1 (CO), 66.5 (OCH2), 48.9 (CHala), 31.6 (OCH2CH2), 28.6 (O[CH2]CH2), 25.6 (CH2CH2CH3), 22.6 (CH2CH3), 15.4 (CH3ala), 13.4 (CH2CH3)
Phenyl-(n-b exyloxy-L-alaninyl)-phosphorochloridate
C15H23N1O4P1Cl1, MW=347.81
This was synthesised according to Standard Procedure 3, using L-Alanine n-hexyl ester hydrochloride salt (0.5 g, 2.38 mmol), PhOP(O)Cl2 (0.36 ml, 2.38 mmol), triethylanine (0.66 ml. 4.77 mmol) in DCM (60 m1). The usual workup yielded the crude product as a yellow oil (0.69 g, 83.2%), which was stored in THF (4 ml) to give a 0.496M solution.
31NMR: xcex49.40, 9.1 0(1:1).
1H NMR: xcex47.44-7.14 (5H,m,OPh), 4.25 (1H,bs,NHala), 4.23-4.03 (3H,m,OCH2+CHala), 1.70-1.63 (2H,m,CH2-2), 1.57-1.54 (2H,m,CH2-3), 1.47-1.32 (7H,m,CH3ala+2CH2-4,5), 0.93-0.91 (3H,dd,CH3-6).
13C NMR: xcex4173.2 (CO), 150.1 (xe2x80x98ipsoxe2x80x99-Ph), 130.3 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 120.9 (xe2x80x98oxe2x80x99-Ph), 66.4 (OCH2), 51.0 (CHala), 31.7 (CH2-C2), 28.9 (CH2-C3), 25.8 (CH2-C4), 22.9 (CH2-C5), 21.0 (CH3ala), 14.4 (CH3-C6).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-Phenyl-(n-hexyloxy-L-alaninyl)-phosphate.
C29H40N7O5P1, MW=597.651
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (100 mg, 0.35 mmol), tBuMgCl (0.7 ml, 0.7 mmol, of a 1.0M solution in THF), in THF (3 ml) and phenyl-(n-hexoxy-L-alaninyl)-phosphorochloridate (2.11 ml, 1.05 mmol, of a 0.496M solution in THF), at room temperature for 24 hrs. Additional phenyl-(n-hexoxy-L-alaninyl)-phosphorochloridate (1.5 ml, 0.68 mmol, of a 0.496M solution in THF), was added and the reaction stirred for a further 24 hrs. The crude product was purified by eluting with 3.0% MeOH in CHCl3 (x2) to give the pure product as a pale yellow foamy solid
31P NMR: xcex43.94, 3.91 (1:1).
1H NMR: xcex47.52 (1H,d,J=8.00 Hz,H8), 7.36-7.31 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.25-7.15 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 6.26 (1H,bs,NHcPr), 6.13-6.08 (1H,m,H2xe2x80x2), 5.93-5.88 (1H,m,H3xe2x80x2), 5.58-5.53 (1H,m,H1xe2x80x2), 5.14 (2H,bs,NH2), 4.28-3.89 (6H,m,CHala,OCH2,H5xe2x80x2+NHala), 3.17 (1H,t,H4xe2x80x2), 3.04 (1H,bs,CHcPr), 2.87-2.75 (1H,m,1of H6xe2x80x2), 1.74-1.61 (3H,m,1of H6xe2x80x2+OCH2CH2), 1.43-1.31 (9H,t,CH3ala+3xCH2), 0.92-0.85 (5H,m,CH3+2Hof CH2cPr), 0.68-0.63 (2H,q,2Hof CH2cPr).
13C NMR: xcex4174.1C0, 160.1 (C2), 156.5 (C4), 151.2 (C6), 151.1 (xe2x80x98ipsoxe2x80x99-Ph), 136.9 (C2xe2x80x2), 136.0 (C8), 131.4 (C3xe2x80x2), 130.0 (xe2x80x98mxe2x80x99-Ph), 125.3 (xe2x80x98pxe2x80x99-Ph), 120.5 (xe2x80x98oxe2x80x99-Ph), 115.0 (C5), 69.2 (C5xe2x80x2), 66.1 (OCH2), 59.3 (Clxe2x80x2), 50.7 (CHala), 46.0 (C4xe2x80x2), 34.9 (C6xe2x80x2), 31.7 (OCH2CH2), 28.8 (CH2-ester), 25.8 (CH2-ester), 24.2 (CHcPr), 21.9 (CH2-ester), 21.5 (CH3ala), 14.4 (CH3-ester), 7.8 (CH2cPr).
L-Alanine cyclo-hexyl ester hydrochloride salt
C9H16N1O2Cl1J, MW=205.71
This was synthesised according to Standard Procedure 2, using L-Alanine (2.0 g, 22.5 mmol), cyclohexanol (2.34 ml, 22.5 mmol), p-toluene sulfonic acid monohydrate (4.7 g, 24.7 mmol), and toluene (100 ml). The p-toluene sulfonate salt was isolated as a pale orange solid (1.45 g).
The reaction was repeated using L-Alanine (3.0 g, 33.7 mmol), cyclohexanol (5.26 ml, 50.6 mmol),p-toluene sulfonic acid monohydrate (9.62 g, 50.6 mmol), and toluene (100 ml). L-alanine cyclohexyl ester hydrochloride salt was isolated as a white solid (3.15 g, 45.45%).
1H NMR (MeOH-d4): xcex44.90 (1Hm,OCH), 4.12-4.04 (1H,q,CHala), 1.92-1.81 (2H,m,OCHCH2), 1.80-1.63 (2H,m,OCHCH2), 1.55 (3H,d,J=7.23 Hz,CH3ala), 1.49-1.33 (6H,m,[CH2]3).
13C NMR: xcex4169.5 (CO), 75.4 (OCH), 48.9 (CHala), 31.3 (2xCH2-o), 25.2 (2xCH2-M), 23.5 (p-CH2), 15.3 (CH3ala).
Phenyl-(c-hexyloxy-L-alaninyl)-phosphorochloridate
C15H21N1O4P1Cl1, MW=345.79
This was synthesised according to Standard Procedure 3, using L-Alanine c-hexyl ester hydrochloride salt (0.7 g, 3.4 mmol), PhOP(O)Cl2 (0.51 ml, 3.4 mmol), triethylamine (0.95 ml 6.8 mmol) in DCM (60 ml). The usual workup yielded the crude product as a yellow oil (1.12 g, 95.2%), which was stored in THF (7 ml) to give a 0.46M solution.
31P NMR: xcex49.43, 9.07 (1:1).
1H NMR: xcex47.44-7.33(2H,m,xe2x80x98oxe2x80x99-Ph), 7.32-7.20 (3Hm,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 4.924.83 (1H,m,OCH), 4.55-4.42 (1H,m,NHala), 4.28-4.15 (1H,m,CHala), 1.89 (2H,bd,CH2-xe2x80x98oxe2x80x99), 1.76 (1H,bd,CH2-xe2x80x98oxe2x80x99), 1.54 (3H,d,CH3ala), 1.49-1.32 (6H,m,CH33CH2-xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99).
13C NMR: xcex4172.5 (CO), 150.1 (xe2x80x98ipsoxe2x80x99-Ph), 130.3 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 121.0 (xe2x80x98oxe2x80x99-Ph), 74.9 (OCH), 51.1 (CHala), 31.8 (CH2-xe2x80x98oxe2x80x99), 25.6 (CH2-xe2x80x98pxe2x80x99), 21.0 (CH3ala).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopeniene-1-methanol O-Phenyl-(c-hexyloxy-L-alaninyl)-phosphate. Cf 1707.
C29H38N7O5P1, MW=595.635
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (100 mg, 0.35 mmol), tBuMgCl (0.7 ml, 0.7 mmol, of a 1.0M solution in THF), in THF (3 ml) and phenyl-(c-hexoxy-L-alaninyl)-phosphorochloridate (2.28 ml, 1.05 mmol, of a 0.46M solution in THF), at room temperature for 24 hrs. The crude product was purified by eluting with 3-4% MeOH in CHCl3, and then 2.5-3.0% MeOH in CHCl3 to give the pure product as a pale yellow foamy solid (199 mg, 95.7%).
31P NMR: xcex44.06, 3.99 (1:1).
1H NMR: xcex47.42 (1H,d,J=8.15 Hz,H8), 7.23-7.18 (2H,n,xe2x80x98oxe2x80x99-Ph), 7.12-7.02 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 6.31 (1H,bs,NHcPr), 5.98 (1H,bs,H2xe2x80x2), 5.78 (1H,bs,H3xe2x80x2), 5.43 (1H,bs,H1xe2x80x2), 5.21 (2H,bs,NH2), 4.66 (1H,bs,OCH), 4.17-4.02 (3H,m,H5xe2x80x2+NHala), 3.95-3.85 (1H,m,CHala), 3.05-2.94 (2H,m,H4xe2x80x2+CHcPr), 2.73-2-63 (1H,m,1of H6xe2x80x2), 1.69 (2H,bs,CH2-xe2x80x98oxe2x80x99), 1.62-1.53 (2H,m,CH2-xe2x80x98oxe2x80x99), 1.45-1.18 (9H,m,CH3ala+3xCH2-xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99xe2x80x2), 0.76 (2H,d,2Hof CH2cPr), 0.53 (2H,bs,2Hof CH2cPr).
13C NMR: xcex4172.OCO, 158.4 (C2), 154.8 (C4), 149.7 (C6), 149.6 (xe2x80x98ipsoxe2x80x99-Ph), 135.5 (C2xe2x80x2), 134.7 (C8), 130.0 (C3xe2x80x2), 128.6 (xe2x80x98mxe2x80x99-Ph), 123.8 (xe2x80x98pxe2x80x99-Ph), 119.1 (xe2x80x98oxe2x80x99-Ph), 113.1 (CS), 72.9 (OCH), 67.8 (C5xe2x80x2), 57.9 (Clxe2x80x2), 59.4 (CHala), 44.7 (C4xe2x80x2), 34.5 (C6xe2x80x2), 30.3 (CH2-xe2x80x98oxe2x80x99), 24.2 (CH2-xe2x80x98mxe2x80x99), 22.8 (CHcPr), 22.5 (CH2-xe2x80x98pxe2x80x99), 20.1 (CH3ala), 6.4 (CH2cPr).
MS ES+: m/z 596.2 (100%, M+), 597.3 (20%, M+H+).
MS FAB: For C29H39O5N7P requires 596.2750, found 596.2750.
HPLC: tR 40.502 (99.8%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
L-alanine cyclobexane-methyl ester hydrochloride
C10H20N1O2Cl1, MW=221.75
This was synthesised according to Standard Procedure 2, using L-Aianine (3.0 g, 33.7 mmol), cyclohexane methanol (4.15 ml, 33.7 mmol), p-toluene sulfonic acid monohydrate (7.05 g, 37.1 mmol), and toluene (100 ml). 9.2 g of the the PTSA salt was solubilised in DCM (50 ml), and washed with 10% K2CO3 (50 ml), and water (2xc3x9750 ml), dried over MgSO4, filtered and the filtrate reduced to dryness to give a yellow oil. This was neutralised with 2M HCl, stirred for 2 hrs, and then freeze-dried to give the hydrochloride salt as a white solid (4.32 g, 75.8%).
1H NMR (MeOH-d4): xcex44.194.01 (3H,m,OCH+CHala), 1.79-1.69 (5H,m,CH+o-CH2), 1.58 (3H,d,J=7.21 Hz,CH3ala), 1.37-1.20 (4H,m,m-CH2), 1.09-0.98 (2H,q,p-CH2).
13C NMR: xcex4170.1 (CO), 71.3 (OCH2), 48.9 (CHala), 37.3 (CH), 29.5 (2xCH2-o), 26.4 (p-CH2), 25.7 (2xCH2-m), 15.4 (CH3ala).
Phenyl-(cyclohexane-methoxy-L-alaninyl)phosphorochloridate
C16H23N1O4P1Cl1, MW=359.82
This was synthesised according to Standard Procedure 3, using L-Alanine cyclohexane-methyl ester hydrochloride salt (0.7 g, 3.16 mmol), PhOP(O)Cl2 (0.47 ml, 3.16 mmol), triethylamine (0.88 ml. 6.31 mmol) in DCM (70 ml). The usual workup yielded the crude product as a yellow oil (1.10 g, 96.8%), which was stored in THF (6 ml) to give a 0.51M solution.
31P NMR: xcex49.35, 9.05 (1:1).
1H NMR: xcex44.614.50 (1H,q,NHala), 4.284.13 (1H,m,CHala), 4.044.00 (2H,q,OCH2), 1.78-1.74 (7H,t,CH2Hx+xe2x80x98oxe2x80x99-CH2), 1.57-1.54 (3H,dd,CH3ala), 1.06-0.96 (2H,q,xe2x80x98pxe2x80x99-CH2).
13C NMR: xcex4173.1 (CO), 150.1 (xe2x80x98ipsoxe2x80x99-Ph), 130.3 (xe2x80x98mxe2x80x99-Ph), 126.4 (xe2x80x98pxe2x80x99-Ph), 121.0 (xe2x80x98oxe2x80x99-Ph), 71.4 (OCH2), 51.0 (CHala), 37.4 (CHcHx), 29.9 (CH2-xe2x80x98oxe2x80x99), 26.7 (CH2-xe2x80x98mxe2x80x99), 25.9 (CH2-xe2x80x98pxe2x80x99), 21.1 (CH3ala).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2cyclopentene-1-methanol O-Phenyl-(cyclobexane-methoxy-L-alaninyl)-phosphate. Cf1708.
C30H40N7O5P1, MW=609.66
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyolopentene-1-methanol (100 mg, 0.35 mmol), tBuMgCl (0.7 mn, 0.7 mmol, of a 1.0M solution in THF), in THF (5 ml) and phenyl-(cyclohexane-methoxy-L-alaninyl)-phosphorochloridate (2.06 ml, 1.05 mmol, of a 0.51M solution in THF), at room temperature for 48 hrs. The crude product was purified by eluting with 4-6% MeOH in DCM, and then 3% MeOH in CHCl3 to give the pure product as a pale yellow foamy solid (161.1 mg, 75.6%).
31p NMR: xcex43.99,3.92 (1:1).
1H NMR: xcex47.40 (1H,d,J=7.07 Hz,H8), 7.24-7.19 (2H,t,xe2x80x98oxe2x80x99-Ph), 7.13-7.03 (3H,m,xe2x80x98mxe2x80x99+xe2x80x98pxe2x80x99-Ph), 6.00-5.96 (2H,m,H2xe2x80x2+NHcPr), 5.79 (1H,qJH3xe2x80x2), 5.45 (1H,d,H1xe2x80x2), 5.05 (2H,bs,NH2), 4.16-4.01 (3H,m,OCH2+NHala), 3.98-3.88 (1H,m,CHala), 3.86-3.74 (2Hm,H5xe2x80x2), 3.07-3.00 (1H,t,H4xe2x80x2), 2.94 (1H,bs,CHcPr), 2.74-2.63 (1H,m,1of H6xe2x80x2), 1.88-1.50. (7H,m,CHcHx+2CH2-xe2x80x98oxe2x80x99), 1.31-1.27 (3H,t,CH3ala), 1.21-0.99 (4H,m,2CH2-xe2x80x98mxe2x80x99), 0.89-0.79 (2H,q,CH2-xe2x80x98pxe2x80x99), 0.75 (2H,d,2Hof CH2cPr), 0.54-0.50 (2H,t,2Hof CH2cPr).
13C NMR: xcex4174.1(CO), 160.2 (C2), 156.4 (C4), 151.2 (C6), 151.1 (xe2x80x98ipsoxe2x80x99-Ph), 136.7 (C2xe2x80x2), 136.0 (C8),131.5 (C3xe2x80x2), 130.0 (xe2x80x98mxe2x80x99-Ph), 125.2 (xe2x80x98pxe2x80x99-Ph), 120.5 (xe2x80x98oxe2x80x99-Ph), 115.1 (C5), 71.0 (OCH2), 69.3 (C5xe2x80x2), 59.3 (Clxe2x80x2), 50.7 (CHala), 46.1 (C4xe2x80x2), 37.4 (CHcHx), 34.9 (C6xe2x80x2), 29.9 (CH2-xe2x80x98oxe2x80x99), 26.6 (CH2-xe2x80x98mxe2x80x99), 25.9 (CH2-xe2x80x98pxe2x80x99), 24.2 (CHcPr), 21.5 (CH3ala), 7.8 (CH2cPr).
MS ES+: miz 610.3 (50%, M+l+), 632.3 (100%, M+Na+), 633.3 (M+H+Na+).
MS FAD: For C30H40O5N7NaP requires 632.2726, found 632.2710.
HPLC: tR 42.859 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
4-Chlorophenyl-phosphorodichloridate
C6H4O2P1Cl3, MW=246.43
Phosphorus oxychloride (2 ml, 21.5mmol) was stirred with anhydrous diethylether (70 ml) in a 250 ml RBF. To this was added, dropwise, a solution of 4-chlorophanol (2.1 ml, 21.5 mmol), and anhydrous triethylamine (3.0 ml, 21.5 mmo) in anhydrous diethylether (30 ml) at xe2x88x9280xc2x0 C. This was stirred vigorously at xe2x88x9280xc2x0 C. for 1 hr and left to rise to room temperature over 16 hrs. The triethylamine hydrochloride salt was filtered off, and the filtrate reduced to dryness to give the crude product as a yellow oil (4.61 g, 87.2%).
3P NMR: xcex44.99 (s).
s3C NMR: xcex4148.4 (xe2x80x98ipsoxe2x80x99-PhX, 133.2 (xe2x80x98pxe2x80x99-Ph), 130.7 (xe2x80x98mxe2x80x99-Ph), 122.4 (xe2x80x98oxe2x80x99-Ph).
4-Chlorophenyl-(methoxy-L-alaninyl)-phosphorochloridate
C10 H12N1O4P1Cl2, MW=246.43
This was synthesised according to Standard Procedure 3, using L-Alanine methyl ester hydrochloride (2.61 g, 18.7 mmol) and p-chlorophenyl phosphorodichloridate (4.61 g, 18.7 mmol) and triethylamine (5.21 ml, 37.4 mmol) in anhydrous DCM (100 ml). The usual workup yielded the crude product as a colourless crude oil (3.76 g, 64.4%) which was stored in anhydrous THF (20 ml) to give a 0.6M solution that was used without further purification.
31 P NMR: xcex49.48, 9.25 (1:1).
1H NMR: xcex47.36 (2H,d,J=8.20 Hz,xe2x80x98oxe2x80x99-Ph), 7.32-7.22 (2H,m,xe2x80x98mxe2x80x99-Ph), 4.69 (1H,d,NHala), 4.274.15 (1H,m,CHala), 3.82 (3H,d,OCH3), 1.56-1.53 (3H,dd,J=7.04 Hz,CH3ala).
13C NMR: xcex4173.4 (CO), 148.6 (xe2x80x98ipsoxe2x80x99-Ph), 131.9 (xe2x80x98pxe2x80x99-Ph), 130.3 (xe2x80x98mxe2x80x99-Ph), 122.3 (xe2x80x98oxe2x80x99-Ph), 53.2 (OCH3), 50.9 (CHala), 20.9 (CH3ala).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[4-chlorophenyl-(methoxy-L-alaninyl)]-phosphate. Cf 1620.
C24H29N7O5P1Cl1, MW=562.02
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (250 mg, 0.87 mmol), tBuMgCl (1.75 ml, 1.75 mmol of a 1.0M solution in THF), and 4-chlorophenyl-(methoxy-L-alaninyl)-phosphorochloridate (4.37 ml, 2.62 mmol, of a 0.6M solution in THF), in anhydrous THF (13 ml) stirring at room temperature for 24 hrs. The crude product was purified by eluting with 3% MeOH in CHCl3 to give the pure product as white foamy solid (364.5 mg, 74.50/%).
31P NMR: xcex44.01 (s).
1H NMR: xcex47.42 (1H,d,HS), 7.22-7.17 (2H,m,xe2x80x98mxe2x80x99-Ph), 7.09-7.03 (2H,t,xe2x80x98oxe2x80x99-Ph), 5.99 (1H,d,H2xe2x80x2), 5.93 (1H,s,H3xe2x80x2), 5.83 (1H,bs,NHcPr), 5.45 (1H,bs,H1xe2x80x2), 4.96 (2H,bs,NH2), 4.11 (2H,bs,H5xe2x80x2), 4.03-3.86 (1H,m,CHala), 3.62 (3H,s,OCH3), 3.07 (1H,d,J=5.9 Hz,H4xe2x80x2), 25 2.92 (1H,bs,CHcPr), 2.76-2.64 (1H,m,1of H6xe2x80x2), 1.64-1.59 (1H,t,1of H6xe2x80x2), 1.32-1.26 (3H,q,CH3ala), 0.76 (2H,d,J=6.40 Hz,2Hof CH2cPr), 0.53 (2H,bs,2Hof CH2cPr).
13C NMR: xcex4174.4 (CO), 160.4 (C2), 156.7 (C4), 151.3 (C6), 149.7 (xe2x80x98ipsoxe2x80x99-Ph), 136.7 (C2xe2x80x2), 135.9 (C8), 131.6 (C3xe2x80x2), 130.5 (xe2x80x98pxe2x80x99-Ph), 130.0 (xe2x80x98mxe2x80x99-Ph), 121.9 (xe2x80x98oxe2x80x99-Ph), 115.2 (C5), 69.4 (C5xe2x80x2), 59.25 (Clxe2x80x2), 52.9(OCH3), 50.6 (CHala), 46.0 (C4xe2x80x2), 34.9 (C6xe2x80x2), 24.1 (CHCPr), 30 21.4 (CH3ala), 7.8 (CH2cPr).
HPLC: tR 32.693, 33.012 (100%)-(100% water (0 mins), 20% water (35rains), 20% water (45 mins), 100% water (55 mins)).
4-Bromophenyl-phosphorodichloridate
C6H4O2Cl2Br1, MW=289.87
This was synthesised by a method analogous to that of 4-chlorophenyl-phosphorodichloridate, except using: Phosphorus oxychloride (3.29 g, 2 ml, 21.5 mmol), and 4-bromophenol (3.71 g, 21.5 mmol) in anhydrous diethylether (70 ml), and anhydrous triethylamine (2.71 g, 3 ml, 21.5 mmol) in anhydrous diethylether (30 ml). The reaction was stirred at xe2x88x9280xc2x0 C. to room temperature for 1 6 hrs. After filtration, and removal of the solvent, the product was obtained as a clear liquid (5.14 g, 82.6%).
31P NMR: xcex44.88 (s).
1H NMR: xcex47.63 (2H,d,J=8.14 Hz,xe2x80x98oxe2x80x99-Ph), 7.28 (2H,t,xe2x80x98mxe2x80x99-Ph),
13C NMR: xcex4149.0(xe2x80x98ipsoxe2x80x99-Ph), 133.7 (xe2x80x98mxe2x80x99-Ph), 122.6 (xe2x80x98oxe2x80x99-Ph), 120.9 (xe2x80x98pxe2x80x99-Ph).
4-Bromophenyl-(methoxy-L-alaninyl)-phosphorochloridate
C10H12N1O4P1Cl1Br1, MW=356.55
This was synthesised according to Standard Procedure 3, using L-alanine methyl ester hydrochloride salt (10 g, 7.1 6 mmol), 4-bromophenyl-phoshorodichloridate (0.82 g, 7.16 mmol), triethylamine (2 ml. 14.3 mmol) in DCM (70 ml). The usual workup yielded the crude product as a yellow oil (2.24 g, 87.7%), which was stored in THF (12 ml) to give a 0.524M solution.
31P NMR: xcex49.16, 9.10 (1:1).
13C NMR: xcex4173.4(CO), 150.1 (xe2x80x98ipsoxe2x80x99-Ph), 133.3 (xe2x80x98mxe2x80x99-Ph), 122.7 (xe2x80x98oxe2x80x99-Ph), 119.6 (xe2x80x98pxe2x80x99-Ph), 53.3 (OCH3), 51.0 (CHala), 20.9 (CH3ala).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[4-bromophenyl-(methoxy-L-alaninyl)]-phosphate. Cf1710.
C24H29N7O5P,Br1, MW=606.42
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (100 mg, 0.35 mmol), tBuMgCl (0.7 ml, 0.7 mmol, of a 1.0M solution in THF), in THF (5 ml) and 4-bromophenyl-(methoxy-L-alaninyl)-phosphorochloridate (2.0 ml, 1.05 mmol, of a 0.524M solution in THF), at room temperature for 24 hrs. The crude product was purified by eluting with 4-6% MeOH in DCM, and then in 4% MeOH in DCM, to give the pure product as a white foamy solid (115.2 mg, 54.4%).
31 p NMR: xcex43.96 (s).
1H NMR: xcex47.42 (1H,d,H8), 7.34-7.30 (2H,dd,J=8.73 Hz,xe2x80x98oxe2x80x99-Ph), 7.03-6.97 (2H,t,J=8.68 Hz,xe2x80x98mxe2x80x99-Ph), 6.02-5.97 (2H,m,H2xe2x80x2+NHcPr), 5.83-5.79 (1H,m,H3xe2x80x2), 5.43 (1H,t,H1xe2x80x2), 5.06 (2H,bs,NH2), 4.28-4.04 (3H,m,H5xe2x80x2+NHala), 4.02-3.85 (1H,m,CHala), 3.61 (3H,d,OCH2), 3.05 (1H,d,J=6.09 Hz,H4xe2x80x2), 2.94 (1H,d,CHcPr), 2.75-2.66 (1H,m,1of H6xe2x80x2), 1.66-1.56 (1Hm,1of H6xe2x80x2), 1.31-1.25 (3H,dd,CH3ala), 0.79-0.72 (2H,q,2Hof CH2cPr), 0.54-0.49 (2H,t,2Hof CH2cPr).
13C NMR: xcex4174.4(CO), 160.3 (C2), 156.6 (C4), 151.3 (C6) 150.2 (xe2x80x98ipsoxe2x80x99-Ph), 136.7 (C2xe2x80x2), 136.0 (C8), 133.0 (xe2x80x98mxe2x80x99-Ph), 131.6 (C3xe2x80x2), 122.4 (xe2x80x98oxe2x80x99-Ph), 118.1 (xe2x80x98pxe2x80x99-Ph), 115.2 (C5), 69.4 (C5xe2x80x2), 59.3 (Clxe2x80x2), 52.9 (OCH3), 50.6 (CHala), 46.0 (C4xe2x80x2), 34.8 (C6xe2x80x2), 24.2 (CHcPr), 21.3 (CH3ala), 7.8 (CH2cPr).
MS ES+: m/z 606.13 (40%, M+), 628.1065 (100%, 79-M+Na+), 630.0967 (95%, 81-M+Na+).
MS FAB: For C24H29O5N7NaPBr requires 628.1049, found 628.1058, and C24H29O5N7NaP
81Br requires 630.1028, found 630.1042.
HPLC: tR 35.882 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
4-Fluorophenyl-phosphorodichlondate
C6H4O2P1Cl2F1, MW=228.97
This was synthesised by a method analogous to that of 4-chlorophenyl-phosphorodichloridate, except using: Phosphorus oxychloride (3.29 g, 2 ml, 21.5 mmol), and 4-fluorophenol (2.41 g, 21.5 mmol) in anhydrous diethylether (70 ml), and anhydrous triethylamine (2.71 g, 3 ml, 21.5 mmol) in anhydrous diethylether (30 ml). The reaction was stirred at xe2x88x9280xc2x0 C. for 4 hrs, and then at room temperature for 2 hrs. After filtration, and removal of the solvent, the product was obtained as a clear liquid (4.08 g, 83.0%).
31P NMR: xcex45.50 (s).
1H NMR: xcex47.29-7.24 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.09 (2H,t,J=8.29 Hz,xe2x80x98mxe2x80x99-Ph), 3C NMR: xcex4159.7(xe2x80x98ipsoxe2x80x99-Ph), 145.8 (xe2x80x98mxe2x80x99-Ph), 122.6 (xe2x80x98oxe2x80x99-Ph), 117.5 (xe2x80x98pxe2x80x99-Ph).
4-Fluorophenyl-(methoxy-L-alaninyl)-phosphorochloridate
C10H12N1O4P1Cl1F1, MW=295.65
This was synthesised according to Standard Procedure 3, using L-alanine methyl ester hydrochloride salt (1.0 g, 7.16 mmol), 4-fluorophenyl-phoshorodichloridate (1.64 g, 7.6 mmol), triethylamine (2 ml. 14.3 mmol) in DCM (70 ml). The usual workup yielded the crude product as a yellow oil (1.97 g, 93.0%), which was stored in THF (12 ml) to give a 0.56M solution.
31P NMR: xcex49.84, 9.60 (1:1).
1H NMR: xcex47.32-7.23(2H,m,xe2x80x98oxe2x80x99-Ph), 7.12-7.06 (2H,m,xe2x80x98mxe2x80x99-Ph), 4.69 (1H,bs,NHala), 4.22 (1H,bs,CHala), 3.82 (3H,d,OCH3), 1.57-1.53 (3H,m,CH3ala).
13C NMR: xcex4173.5(CO), 161.6 (xe2x80x98ipsoxe2x80x99-Ph), 145.9 (xe2x80x98mxe2x80x99-Ph), 122.5 (xe2x80x98oxe2x80x99-Ph), 117.0 (xe2x80x98pxe2x80x99-Ph), 53.2 (OCH3), 50.9 ((Hala), 20.9 (CH3ala).
(1S,4R4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[(4-fluoropheny]-(methoxy-L-alaninyl)]-phosphate. Cf1737.
C24H29N7O5P1F1, MW=545.57
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (100 mg, 0.35 mmol), tBuMgCl (0.7 ml, 0.7 mmol, of a 1.0M solution in THF), in THF (5 ml) and 4-fluorophenyl-(methoxy-L-alaninyl)-phosphorochloridate (1.89 ml, 1.05 mmol, of a 0.56M solution in THF), at room temperature for 24 hrs. The solvent was removed under reduced pressure and the residue columned in 2.5-5% methanol in chloroform, and then in 3% methanol in chloroform, to give the pure product as a pale yellow foamy solid (62.0 mg, 32.5%).
31 P NMR: xcex44.24, 4.23, 4.20, 4.19.
1H NMR: xcex47.52 (1H,d,H8), 7.21-7.14 (2H,m,xe2x80x98oxe2x80x99-Ph), 7.03-6.97 (2H,m,xe2x80x98mxe2x80x99-Ph), 6.16 (1H,bs,NHcPr), 6.10-6.07 (1H,q,H2xe2x80x2), 5.93-5.89 (1H,q,H3xe2x80x2), 5.44 (1H,d,H1xe2x80x2), 5.14 (2H,bs,NH2), 4.23-3.98 (4H,m,H5xe2x80x2,NHala+CHala), 3.72 (3H,d,OCH2), 3.16 (1H,d,J=6.03 Hz,H4xe2x80x2), 3.03 (1H,d,CHcPr), 2.86-2.74 (1H,m,1of H6xe2x80x2), 1.76-1.66 (1H,m,1of H6xe2x80x2), 1.42-1.35 (3H,dd,CH3ala), 0.89-0.83 (2H,q,2Hof CH2cPr), 0.65-0.60 (2H,t,2Hof CH2cPr).
13C NMR: xcex4174.4(CO), 161.5 (C2), 160.3+156.6 (xe2x80x98pxe2x80x99-Ph), 156.6 (C4), 151.3 (C6) 150.2 (xe2x80x98ipsoxe2x80x99-Ph), 136.8 (C2xe2x80x2), 136.0 (C8), 131.6 (C3xe2x80x2), 121.9 (xe2x80x98oxe2x80x99-Ph), 115.1 (C5), 69.3 (C5xe2x80x2), 59.3 (Clxe2x80x2), 52.9 (OCH3), 50.6 (CHala), 46.0 (C4xe2x80x2), 34.9 (C6xe2x80x2), 24.2 (CHcPr), 21.3 (CH3ala), 7.8 (CH2cPr).
HPLC: tR 31.536 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
4-Iodophenyl-phosphorodichloridate
C6H4O2P1Cl2I1, MW=336.07
This was synthesised by a method analogous to that of 4-chlorophenyl-phosphorodichloridate, except using: Phosphorus oxychloride (3.29 g, 2 ml, 21.5 mmol), and 4-iodophenol (4.72 g, 21.5 mmol) in anhydrous diethylether (60 ml), and anhydrous triethylamine (2.71 g, 3 ml, 21.5 mmol) in anhydrous diethylether (20 ml). The reaction was stirred at xe2x88x9280xc2x0 C. for 4 hrs, and then at room temperature for 2 hrs. After filtration, and removal of the solvent, the product was obtained as a clear liquid (6.2 g, 85.8%).
31P NMR: xcex44.72 (s).
1H NMR: xcex47.71 (2H,d,J=8.59 Hz,xe2x80x98oxe2x80x99-Ph), 7.06-7.02 (2H,dd,J=8.80 Hz,xe2x80x98mxe2x80x99-Ph),
1C NMR: xcex4149.9(xe2x80x98ipsoxe2x80x99-Ph), 139.8 (xe2x80x98mxe2x80x99-Ph), 122.9 (xe2x80x98oxe2x80x99-Ph), 91.9 (xe2x80x98pxe2x80x99-Ph).
4-Iodophenyl-(methoxy-L21alaninyl)-phosphorochloridate
C10H12N1O4P1Cl1,I1, MW=403.55
This was synthesised according to Standard Procedure 3, using L-alanine methyl ester hydrochloride salt (1.0 g, 7.16 mmol), 4-iodophenyl-phoshorodichloridate (2.41 g, 7.16 mmol), triethylamine (2 ml. 14.3 mmol) in DCM (70 ml). The usual workup yielded the crude product as a yellow oil (3.59 g,  greater than 100%), which was stored in THF (14 ml) to give a 0.51M solution.
31P NMR. xcex49.31, 9.08 (1:1).
1H NMR: 7.74-7.69(2H,m,xe2x80x98oxe2x80x99-Ph), 7.32-7.05 (2H,m,xe2x80x98mxe2x80x99-Ph), 4.73 (1H,bs,NHala), 4.20 (1H,bs,CHala), 3.81 (3H,d,0CH3), 1.56-1.53 (3H,dd,J=7.06 Hz,CH3ala).
13C NMR: xcex4173.4(CO), 149.9 (xe2x80x98ipsoxe2x80x99-Ph), 139.5 (xe2x80x98mxe2x80x99-Ph), 123.0 (xe2x80x98oxe2x80x99-Ph), 90.4 (xe2x80x98pxe2x80x99-Ph), 53.3 (OCH3), 50.9 (CHala), 20.9 (CH3ala).
(1S,4R)-4-(2-amino6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[4-iodophenyl-(methoxy-L-alaninyl)]-phosphate. Cf1738.
C24H29N7O5P1I1, MW=653.48
This was synthesised according to Standard Procedure 4, using (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (100 mg, 0.35 mmol), tBuMgCl (0.7 ml, 0.7 mmol, of a 1.0M solution in THF), in THF (5 ml) and 4-iodophenyl-(methoxy-L-alaninyl)-phosphorochloridate (2.05 ml, 1.05 mmol, of a 0.51M solution in THF), at room temperature for 48 hrs. The solvent was removed under reduced pressure and the residue columned in 3-6% methanol in chloroform, and then in 3% methanol in chloroform, to give the pure product as a white foamy solid (82.0 mg, 29.9%).
31 P NMR: xcex43.92 (s).
1H NMR: xcex47.63-7.59 (2H,dd,J=8-65 Hz,xe2x80x98mxe2x80x99-Ph), 6.98 (2H,t,J-8.20 Hzxe2x80x98oxe2x80x99-Ph), 6.25 (1H,bs,NHcPr), 6.09 (1H,t,H2xe2x80x2), 5.91 (1H,t,H3xe2x80x2), 5.54 (1H,d,H1xe2x80x2), 5.21 (2H,bs,NH2), 4.35-4.16 (3H,m,H5xe2x80x2,NHala), 4.07-3.95 (1H,m,CHala), 3.71 (3H,d,OCH3ala), 3.15 (1H,d,J=7.23 Hz,H4xe2x80x2), 3.03 (1H,bs,CHcPr), 2.85-2.74 (1H,m,1of H6xe2x80x2), 1.761.65 (1H,m,1of H6xe2x80x2), 1.43-1.35 (3H,t,CH3ala), 0.89-0.83 (2H,q,2Hof CH2cPr), 0.63 (2H,bs,2Hof CH2cPr).
13C NMR: xcex4174.4(CO) 160.2 (C2), 156.5 (C4), 151.1 (C6) 151.0 (xe2x80x98ipsoxe2x80x99-Ph), 139.0 (C2xe2x80x2), 136.8 (xe2x80x98mxe2x80x99-Ph), 136.0 (C8), 131.5 (C3xe2x80x2), 122.8 (xe2x80x98oxe2x80x99-Ph), 115.0 (C5), 88.9 (xe2x80x98pxe2x80x99-Ph), 69.4 (C5xe2x80x2), 59.3 (C1xe2x80x2), 52.9 (OCH3), 50.6 (CHala), 46.0 (C4xe2x80x2), 34.8 (C6xe2x80x2), 24.2 (CHcPr), 21.3 (CH3ala), 7.8 (CH2cPr).
HPLC: tR 33.848 (100%)-(100% water (0 mins), 20% water (35 mins), 20% water (45 mins), 100% water (55 mins)).
L-Alanine (3-pentyl) ester hydrochloride salt 
Tionyl chloride (1.6 ml, 0.022 M) was added dropwise to a stirred solution of 3-pentanol (18.2 ml, 0.17 M) at 0xc2x0 C. under nitrogen. The mixture was stirred for 30 minutes, then allowed to warm to room temperature. L-Alanine (pre-dried at 60xc2x0 C. over P2O5 for 4 hrs: 1.0 g, 0.011 M) was added and the resulting suspension was heated at reflux overnight (the reaction mixture became a clear, colourless solution). The solvent was removed under reduced pressure to leave an oil which was repeatedly triturated and coevaporated with diethyl ether, then petrol (60/80) to remove traces of 3-pentanol. The resulting oily residue solidified on drying under high vacuum to give a peach-coloured solid (1.96 g, 10 mmol, 89%).
xcex4H (d4-CH3OH, 300 MHz) 0.94 (t, 6H, O-CH(CH2CH3)2, J =7), 1.57 (d, 3H, CH3-ala, J=7), 1.67 (m, 4H, O-CH(CH2CH3)2, J=7), 4.12 (q, 1H, CH-ala, J =7), 4.88 [m, 1H, O-CH(C2H2)2]; xcex4C(d4-CH3OH, 75 MHz) 8.87 [O-CH(CH2CH3)2], 15.38 (CH3-ala), 26.39, 26.44 [O-CH(CH2CH3)2J, 48.82 (CH-ala), 79.88 [O-CH(C2H)2], 170.03 (Cxe2x95x90O).
Phenyl (3-pentyloxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.45 ml, 3.0 mmol), dry triethylamine (0.8 ml, 6.0 mmol), L-alanine (3-pentyl) ester hydrochloride salt 1a (0.583 g, 3.0 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, pale yellow oil (1.055 g,  greater than 100%)
xcex4P (CDCl3, 121 Mz) 8.99, 9.37
The product was redissolved in dry THF 5 ml) and used as a 0.211 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl (3-pentyloxy-L-alaninyl)phosphate [Cf 1685]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol), phenyl(3-pentyloxy-L-alaninyl)phosphorochloridate 1b (3.3 ml of 0.211 g/ml solution 4 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 1.5 hrs. The crude residue was purified twice by column chromatography, using (i) MeOH:CHCl3 (4:96) and (ii) MeOH:CHCl3 (3:97) as eluent, to give the product as a clear, colourless oil, which solidified to a white foam after trituration and coevaporation with diethyl ether (0.202 g, 0.35 mmol, 50%). xcex4P (CDCl3, 121 MHz) 3.89; 31 (CDCl3, 300 MHz) 0.66 (m, 2H, CH2-CPr), 0.90 [m, 8H CH2-cPr and CH(CH2CH3)2], 1.43 (m, 3H, CH3-ala), 1.58 [m, 4H, CH(CH2CH3)2], 1.72 (m, 1H, 61H.), 2.82 (m, 1H, 6xe2x80x2Hb), 3.05 (m, 1H, C1H-cPr), 3.20 (m, 1H, 4xe2x80x2H), 3.77 (m, 1H, CH-ala), 4.05 (m, 1H, NH-ala), 4.22 (m, 2H, 51H), 4.80 (m, 1H, O-CHxe2x80x94), 4.89 (s, 2 M, NH2), 5.56 (m, 1H, 1xe2x80x2H), 5.78 (bs, 1H, NH-cPr), 5.93 (m, 1H, 31H), 6.12 (m, 1H, 21H), 7.26 (m, 5H, ArH), 7.51 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 6.37 (CH2-cPr), 8.50 [CH(CH2CH3)2], 20.28 (CH3-ala), 22.68 (CH-cPr), 25.28, 25.38 [CH(CH2CH3)2], 33.51, 33.60 (6xc2x0 C.), 44.59, 44.69 (4xc2x0 C.), 49.40 (CH-ala), 57.79, 57.83 (1xc2x0 C.), 67.90 (5xc2x0 C.), 7729 (OCH), 113.86 (5C), 119.10-119.18 (o-Ph), 123.84 (p-Ph), 128.61 (m-Ph), 130.09, 130.16 (3xe2x80x2C), 134.45, 134.56 (8C), 135.27, 135.41 (2xe2x80x2C), 149.66-149.93 (6C and ipso-Ph), 155.26 (4C), 158.95 (2C), 172.32, 172.44 (Cxe2x95x90O); m/z (FAB) 584.2751 (MW, C28H39N7O5P requires 584.2750).
L-Alanine (3,3-dimethyl-1-butyl) ester hydrochloride salt 
Prepared according to Standard Procedure 2, from L-alanine (1.6 g, 18 mmol), PTSA monohydrate (3.8 g, 20 mmol), 3,3-dimethyl butan-1-ol (2.2 ml, 18 mmol) and toluene (100 ml). Conversion to the hydrochloride salt: the p-toluene sulfonate salt was redissolved in CHCl3 and washed with 10% potassium carbonate solution and water. The organuc layer was dried (MgSO4), filtered and the solvent was removed under reduced pressure to give the crude product as an oil. Aq. HCl (1 M), was added and the solution stirred for 30 minutes at room temperature. The solution was freeze-dried to give the hydrochloride salt as a white solid (3.31 g, 15.8 mmol, 88%).
xcex4H (d4-CH3OH, 300M ) 0.93 [s, 9H, Oxe2x80x94(CH2)2(CH3)3], 1.50 (d, 3H, CH3-ala, J=7), 1.59 (t, 2H, O-CH2CH2, J=7), 4.05 (q, 1I, CH-ala, J 7), 4.25 (m, 2H, Oxe2x80x94CH2); 8c (d4-CH3OH, 75 MHz) 15.18 (CH3-ala), 28.91 [C(CH3)3], 29.54 [C(CH3)3], 41.62 (Oxe2x80x94CH2CH2xe2x80x94), 48.85 (CH-ala), 64.11 ()xe2x80x94CH2CH2xe2x80x94), 170.03 (Cxe2x95x90O).
Phenyl(3,3-dimethyl-1-butoxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.45 ml, 3.0 mmol), dry triethylamine (0.8 ml, 6.0 mmol), L-alanine (3,3-dimethyl-1-butyl) ester hydro-chloride salt 2a (0.632 g, 3.0 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, pale yellow oil (1.038 g, 99%).
xcex4p (CDCl3, 121 MHz) 8.94, 9.30
The product was redissolved in dry THF (5 ml) and used as a 0.208 g/ml solution
(1S,4R)-4-(2-amino6cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(3,3-dimethyl-]-butoxy-L-alaninyl)phosphate [Cf 1687]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol), phenyl (3,3-dimethyl-1-butoxy-L-alaninyl) phosphorochloridate 2b (3.5 ml of 0.208 g/ml solution, 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 1.5 hrs. The crude residue was purified twice by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to a white foam after trituration and coevaporation with diethyl ether (0.287 g, 0.5 mmol, 69%). 5? (CDCl3, 121 M) 3.83; 5H (CDCl3, 300 M) 0.66 (m, 2H, CH2,-cPr), 0.90 (m, 2H, CH2cPr), 0.97 [s, 9H, C(CH3)3]1.41 (m, 3H1, CH3-ala), 1.57 (m, 2H, Oxe2x80x94CH2CH2), 1.74 (m, 111, 61H.), 2.82 (m, 111, 6xe2x80x2Hb), 3.05 (m, 1ill CH-cPr), 3.20 (m, 1H, 4xe2x80x2H), 3.70 (m, 1H CH-ala), 4.04 (m, 1H, NH-ala), 4.22 (i, 4H, 5xe2x80x2H and C-CH2CH2), 4.88 (s, 2H, NH2), 5.57 (m, 1H, 1xe2x80x2H), 5.75 (bs, 1H, NH-cPr), 5.93 (m, 1H, 3xe2x80x2H), 6.12 (m, 1H, 2xe2x80x2H), 7.27 (m, 5H, ArH), 7.52 (d, 1H, 8H); 5c (CDCl3, 75 MHz) 6.35 (CH2-cPr), 19.95, 20.01 (CH3-ala), 22.69 (CH-cPr), 28.52 [C(CH3)3], 28.52 [C(CH3)3], 33.49, 33.57 (6xe2x80x2C), 40.59, 40.63 (OCH2CH2xe2x80x94), 44.58, 44.68 (4xe2x80x2C), 49.28 (CH-ala), 57.79, 57.83 (1xe2x80x2C), 62.28, 62.31 (OCH2CH2xe2x80x94), 67.86, 67.94 (5xe2x80x2C), 113.81 (5C), 119.10, 119.16 (p-Ph), 123.84 (o-Ph), 128.61 (m-Ph), 130.10, 130.16 (3xe2x80x2C), 134.47,134.56 (8C), 135.29, 135.40 (2xe2x80x2C), 149.67-149.75 (6C and ipso-Ph), 155.25 (4C), 158.96 (2C), 172.55, 172.65 (Cxe2x95x90O); m/z (FAB) 598.2896 (MH+, C29H41 N7O5P requires 598.2907).
L-Alanine (4-methyl-1-pentyl) ester p-toluene sulfonate salt 
Prepared according to Standard Procedure 2, from L-alanine (1.6 g, 18 mmol), p-TSA monohydrate (3.8 g, 20 mmol), 4-methyl pentan-1-ol (2.24 ml, 18 mmol) and toluene (100 ml). The p-toluene sulfonate salt was isolated as a white solid (6.082 g, 17.6 mmol, 98%). 8H (d4-CH30H, 300 MHz) 0.93 [d, 6H, CH(CH3)2], 127 (m, 21I, O-CH2CH2CH2xe2x80x94), 1.54 (d, 3H, CH3-ala), 1.59 (m, 1H, CH(CH3)2), 1.69 [m, 2H, Oxe2x80x94(CH2)2C, 2.39 (s, 3H, CH3, p-TSA), 4.10 (m, 1H, CH-ala), 4.24 (m, 2H, O-CH2), 7.25 (d, 2H, ArH, p-TSA), 7.72 (d, 2H, ArH, p-TSA); xcex4C (d4-CH3OH, 75 MHz) 15.23 (CH3-ala), 20.31 (CH3-pTSA), 21.83 [CH(CH3)2], 26.45 (O-CH2CH2CH2xe2x80x94), 27.87 [CH(CH3)2], 34.93 (Oxe2x80x94CH2CH2CH2xe2x80x94), 48.85 (CH-ala), 66.77 [Oxe2x80x94CH2(CH2)2], 125.93 (o-Ph, p-TSA), 128.83 (m-Ph, p-TSA), 140.75 (ipso-C-CH3,p-TSA), 142.39 (ipso-C-S, p-TSA), 170.07 (Cxe2x95x90O).
Phenyl(4-methyl-1-pentyloxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.9 ml, 6.0 mmol), dry triethylamine (1.7 ml, 12.0 mmol), L-alanine (4-methyl-1-pentyl) ester p-toluene sulfonate salt 3a (2.081 g, 6.0 mmol) and dry DCM (100 ml total). The crude product was obtained as a clear, colourless oil (1.79 g, 85%).
xcex4P (CDCl3, 121 MHz) 8.95, 9.31
The product was redissolved in dry THF (10 ml) and used as a 0.179 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-y))-2-cyclopentene-1-methanol O-[phenyl (4-methyl-1-pentyloxy-L-alaninyl)phosphate [Cf 1721]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol), phenyl(4-methyl-1-pentyloxy-L-alaninyl)phosphorochloridate 3b (4.1 ml of 0.179 g/ml solution, 2.1 mmol) and dry THF (10 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 3 hrs. The crude residue was purified by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to a white foam after trituration and coevaporation with diethyl ether (0.288 g, 0.5 mmol, 69%).
xcex4P (CDCl3, 121 M) 3.84, 3.88; xcex4H (CDCl3, 300 M) 0.64 (m, 2H, CH2-CPr), 0.87 (m, 2H, CH2CPr), 1.24 [m, 2H, CH(CH3),2], 1.40 (t, 3H, CH3-ala), 1.60 [m, 3H, CH(Cl3)CH3], 1.73 [m, 3H, CH(CH3)CH3], 2.19 (m, 1H, 6xe2x80x2Hb), 2.80 (m, 1H, 6xe2x80x2Hb), 3.03 (m, 1H, CHCPr), 3.18 (m, 1H, 4xe2x80x2H), 3.88 (m, 1H, CH-ala), 4.03 (m, 3H, OCH2xe2x80x94 and NHxe2x80x94ala), 4.21 (m, 2H, 5xe2x80x2H), 4.99 (bs, 2H, NH2), 5.55 (m, 1xe2x80x2H, 1xe2x80x2H), 5.91 (m, 2H, NH-cPr and 31H), 6.10 (m, 1H, 21H), 7.29 (m, 5H, ArH), 7.51 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 7.79 (CH2-cPr), 21.55 (CH3-ala), 21.61 [CH(CH3)2], 23.69 (CH-cPr), 25.66 (Oxe2x80x94CH2CH2CH2xe2x80x94), 29.63 [CH(CH3)2], 35.00 (6xe2x80x2C), 38.81 (O-CH2CH2CH2xe2x80x94), 46.01, 46.11 (4xc2x0 C.), 50.72 (CH-ala), 59.21 (1xe2x80x2C), 69.31 (5xe2x80x2C), 69.85 (O-CH2CH2-), 115.25 (5C), 120.52-120.62 (p-Ph), 125.25 (o-Ph), 130.04 (m-Ph), 131.59 (3xe2x80x2C), 135.98 (8C), 136.71, 136.79 (2xe2x80x2C.), 151.08, 151.17 (6C and ipso-Ph), 156.70 (4C), 160.40 (2C), 174.00, 174.10 (Cxe2x95x90O); m/z (FAB) 598.2883 (MH+, C29H41N7O5P requires 598.2907).
L-Alanine (cyclopropyl methyl) ester hydrochloride salt 
Thionyl chloride (1.2 ml, 0.017 M) was added dropwise to a stirred solution of cyclopropyl methanol (6.8 ml, 8.4 mmol) at 0xc2x0 C. under nitrogen. The mixture was stirred for 30 minutes, then allowed to warm to room temperature. L-Alanine (pre-dried at 60xc2x0 C. over P2O5 for 4 hrs: 0.75 g, 8.4 mmol) was added and the resulting suspension was heated at reflux overnight (the reaction mixture became a clear, colourless solution). The solvent was removed under reduced pressure to leave an orange/red oil which was repeatedly triturated and coevaporated with diethyl ether, to remove traces of cyclopropyl methanol. Diethyl ether (200 ml) was added and the mixture was stirred for 30 min. The resulting suspension was filtered to give the product as a cream solid (1.29 g, 7.1 mmol, 85%).
xcex4H(d4-C:H3OH, 300 MHz) 0.38 (m, 21, CH2-cPr), 0.65 (m, 2H, CH2-cPr), 1.24 (m, 1H, CH-cPr), 1.60 (d, 3H, CH3-ala, J=7), 4.13 (m, 3H, CH-ala and O-CH2); xcex4C (d4-CH3OH, 75 MHz) 4.17 (CH2-cPr), 10.98 (CH-cPr), 16.72 (CH3-ala), 50.33 (CH-ala), 72.70 (O-CH2), 171.56 (Cxe2x95x90O).
Phenyl(cyclopropyl methoxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.9 ml, 6.0 mmol), dry triethylamine (1.7 ml, 12.0 mmol), L-alanine (cyclopropyl methyl) ester p-toluene sulfonate salt 4a (1.082 g, 6.0 mmol) and dry DCM (100 ml total). The crude product was obtained as a clear, yellow oil (1.79 g, 94%).
xcex4P(CDCl3, 121) 9.00, 9.36
The product was redissolved in dry THF (5 ml) and used as a 0.385 g/ml solution.
(1S,4R)-4(2-amino6-cyclopropylamino-9H-purin-9-yl)-2-cyclopeutene-1-methanol O-[phenyl(cyclopropyl methoxy-L-alaninyl)]phosphate 
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol) phenyl(cyclopropyl methoxy-L-alaninyl) phosphorochloridate 4b (1.85 ml of 0.385 g/ml solution, 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 3.5 hrs. The crude residue was purified twice by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil which solidified to a white foam after trituration and coevaporation with diethyl ether (0.244 g, 0.4 mmol, 61%).
xcex4P (CDCl3, 121 MHz) 3.88, 3.94; xcex4H (CDCl3, 300 MHz) 0.29 (m, 2H, CH2-cPr), 0.61 (m, 2H, CH2-cPr), 0.87 (m, 2H, CH2-cPr), 1.17 (m, 1H, CH-cPr), 1.42 (t, 3H, CH3-ala), 1.69 (m, 1H, 6xe2x80x2Ha), 2.81 (m, 1H, 6xe2x80x2Hb), 3.04 (m, 1H, CH-cPr), 3.18 (m, 1H, 4xe2x80x2H), 4.01 (m, 41, OCH2xe2x80x94, CH-ala and NH-ala), 4.21 (m, 2H, 51H), 5.03 (bs, 2H, NH2), 5.56 (m, 1I, 1xe2x80x2H), 5.91 (m, 1H, 31H), 6.05 (bs, 1H, NH-cPr), 6.10 (m, 1H, 21H), 7.25 (m, 5H, ArH), 7.51 (d, 111H, 8H); 5c (CDCl3, 75 MHz) 2.24 (CH2-cPr), 6.33 (CH2-cPr), 8.66 (CH2-cPr), 20.05, 20.11 (CH3-ala), 22.68 (CH-cPr), 33.55 (6xe2x80x2C), 44.58, 44.68 (4xe2x80x2C), 49.24, 49.31 (CH-ala), 57.77, 57.82 (1xe2x80x2C), 67.76, 67.93 (0-CH2), 69.27, 69.29 (5xe2x80x2C), 113.74 (5C), 119.10-119.19 (p-Ph), 123.84 (o-Ph), 128.61 (m-Ph), 130.09, 130.13 (3xe2x80x2C), 134.42, 134.50 (8C), 135.32, 135.40 (2xe2x80x2C), 149.66, 149.74 (6C and ipso-Ph), 155.26 (4C), 159.00 (2C), 172.64, 172.73 (Cxe2x95x90O).
L-Alanine (cyclobutyl methyl) ester hydrochloride salt 
Prepared according to Standard Procedure 2, from L-alanine (1.6 g, 18 mmol), p-TSA monohydrate (3.8 g, 20 mmol), cyclobutane methanol (1.9 ml, 20 mmol) and toluene (100 ml). The p-toluene sulfonate salt was isolated as a white solid (4.249 g, 12.9 mmol, 72%).
xcex4H (d4-CH30H, 300 MHz) 1.54 (d, 3H, CH3-ala, J=7), 1.89 (m, 4H, cBu-2/4H), 2.08 (m, 2H, cBu-3H), 2.39 (s, 3H, CH3, p-TSA), 2.69 9 m, 1H, CH-cBu), 4.11 (q, 11, CH-ala, J=7), 4.22 (m, 2H, Oxe2x80x94CH2), 7.26 (d, 2H, ArH, p-TSA), 7.73 (d, 2H, ArH, p-TSA); xcex4C (d4-CH3OH, 75 MHz) 16.7 (CH3-ala), 19.6 (CH2-cBu), 21.7 (CH3-pTSA), 25.9 (CH2-cBu), 35.7 (CH-cBu), 48.9 (CH-ala), 71.3 (Oxe2x80x94CH2), 127.4 (o-Ph, p-TSA), 130.3 (m-Ph, p-TSA), 142.2 (ipso-C-CH3,p-TSA), 143.8 (ipso-C-S, p-TSA), 171.6 (Cxe2x95x90O).
Phenyl(cyclobutyl methoxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.9 ml, 6.0 1 mmol), dry triethylamine (1.7 ml, 12.0 mmol), L-alanine (cyclobutyl methyl) ester p-toluene sulfonate salt 5a (1.98 g, 6.0 mmol) and dry DCM (100 ml total). The crude product was obtained as a clear, colourless oil (2.04 g,  greater than 100%).
xcex4P (CDCl3, 121 ) 9.00, 9.34
The product was redissolved in dry THF (5 ml) and used as a 0.408 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(cyclobutyl methoxy-L-alaninyl)]phosphate [Cf 1773]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in TEF: 1.4 ml, 1.4 mmol) phenyl(cyclobutyl methoxy-L-alaninyl) phosphorochloridate 5b (1.7 ml of 0.408 g/ml solution, 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 3 hrs. The crude residue was purified twice by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to a white foam after trituration and coevaporation with diethyl ether (0.213 g, 0.4 mmol, 52%).
xcex4P (CDCl3, 121M ) 3.87, 3.91; 3H (CDCl3, 300M ) 0.65 (m, 2H, CH2cPr), 0.89 (m, 2H, CH2-cPr), 1.41 (t, 3H, CH3-ala), 1.74 (m, 3H, CH2-cBu and 61H,), 2.06 (m, 2H, CH2-cBu), 2.61 (m, 2H, CH2-cBu), 2.81 (m, 1H, 61Hb), 3.04 (m, 1H, CH-cPr), 3.19 (m, 1H, 4xe2x80x2H), 3.90 (m, 1H, NH-ala), 4.09 (m, 3H, OCH2xe2x80x94, and CH-ala), 4.22 (m, 2H, 5xe2x80x2H), 4.98 (bs, 2H, NH2), 5.56 (m, 1H, 11H), 5.92 (m, 2H, 31H and NH-cPr), 6.11 (m, 1H, 2xe2x80x2H), 7.26 (m, 5H, ArH), 7.52 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 6.37 (CH2-cPr), 17.33 (CH2-cBu), 20.17, 20.23 (CH3-ala), 22.68 (CH-cPr), 23.57 (2xc3x97CH2-cBu), 32.86 (CH-cBu), 33.51, 33.55 (6xe2x80x2C), 44.58, 44.68 (4xe2x80x2C), 49.23, 49.28 (CH-ala), 57.81, 57.85 (1xe2x80x2C), 67.78-67.94 (5xe2x80x2C), 68.17, 68.20 (O-CH2), 113.83 (5C), 119.09-119.19 p-Ph), 123.87 (o-Ph), 128.62 (m-Ph), 130.11, 130.15 (3xe2x80x2C), 134.51, 134.61 (8C), 135.30,135.39 (2xe2x80x2C), 149.64149.97 (6C and ipso-Ph), 155.20 (4C), 158.87 (2C), 172.64, 172.74 (Cxe2x95x90O).
L-Alanine (cyclopentyl methyl) ester p-toluene sulfonate salt 
Prepared according to Standard Procedure 2, from L-alanine (1.6 g, 18 mmol), p-TSA monohydrate (3.8 g, 20 mmol), cyclopentane methanol (1.9 ml, 18 mmol) and toluene (100 ml). The p-toluene sulfonate salt was isolated as a white solid (6.21 g, 18 mmol, 100%). 5H (d4-CH3OH, 300 MH:) 1.22 (m, 2H, cPent 2/5H.), 1.46 (d, 3H, CH3-ala), 1.56 (m, 4H, cPent 2/3/4/5Hb), 1.70 (m, 2H, cPent 3/4H.), 2.19 (m, 1xe2x80x2H, CH-cPent), 2.31 (s, 3H, CH3, p-TSA), 4.06 (m, 3H, O-CH2 and CH-ala), 7.18 (d, 2H, ArH, p-TSA), 7.64 (d, 2H, ArH, p-TSA);
67xcex4C (d4-CH3OH, 75 MHz) 15.25 (CH3-ala), 20.30 (CH3,p-TSA), 25.27 (CH2-cPent), 29.10, 29.15 (CH2-cPent), 38.72 (CH-cPent), 48.84 (CH-ala), 70.12 (O-CH2), 125.93 (o-Ph, p-TSA), 128.82 (m-Ph, p-TSA), 140.75 (ipso-C-CH3, p-TSA), 142.40 (ipso-C-S, p-TSA), 170.09 (Cxe2x95x90O).
Phenyl(cyclopentyl methoxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.9 ml, 6.0 mmol), dry triethylamine (1.7 ml, 12.0 mmol), L-alanine (cyclopentane methyl) ester p-toluene sulfonate salt 6a (2.069 g, 6.0 mmol) and dry DCM (100 ml total). The crude product was obtained as a clear, yellow oil (1.97 g, 95%).
xcex4P (CDCl3, 121 M)8.94, 9.30
The product was redissolved in dry THF (IO ml) and used as a 0.197 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(cyclopentyl methoxy-L-alaninyl)phosphate [Cf 1722]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol), phenyl (cyclopentene methoxy-L-alaninyl) phosphorochloridate 6b (3.7 ml of 0.197 g/ml solution, 2.1 mmol) and dry THF (10 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 3 hrs. The crude residue was purified by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to a white foam after trituration and coevaporation with diethyl ether (0.314 g, 0.5 mmol, 75%).
xcex4P (CDCl3, 121 MHz) 3.86, 3.87; 5H (CDCl3, 300 MHz) 0.65 (m, 2H, CH2-cPr), 0.89 [m, 8H, CH2-cPr and (CH2)3-cPent], 1.24 (m, 2H, CH2-cPent), 1.41 (m, 311, CH3-ala), 1.65 (m, 2H, CH-cPent and 61H.), 2.81 (m, 1H, 6xe2x80x2Hb), 3.04 (in, 1H, CH-cPr), 3.19 (m, 1H, 41H), 3.80 (in, 1H, CH-ala), 4.07 (m, 3H, OCH2 and NH-ala), 4.22 (m, 2H, 5xe2x80x2H), 4.92 (bs, 2H, NH2), 5.55 (m, 1H, 1xe2x80x2H), 5.81 (bs, 1H, NH-cPr), 5.92 (in, 1H, 31H), 6.11 (m, 1H, 2xe2x80x2H), 7.26 (m, 5H, ArH), 7.52 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 6.42 (CH2-cPr), 21.43 (CH3-ala), 22.73 (CH-cPr), 24.59 (CH2-cPent), 25.35 (CH2-cPent), 26.64 (CH-cPent), 33.43, 33.51 (6xe2x80x2C), 44.58, 44.68 (4xe2x80x2C), 49.23 (CH-ala), 57.86, 57.91 (1xe2x80x2C), 64.69, 64.97 (O-CH2xe2x80x94), 67.84 (5xe2x80x2C), 113.74 (5C), 119.09-119.18 (p-Ph), 123.88 (o-Ph), 128.62 (m-Ph), 130.05, 130.11 (3xe2x80x2C), 134.65,134.76 (8C), 135.33, 135.44 (2xe2x80x2C), 149.63, 149.72 (6C and ipso-Ph), 154.98 (4C), 158.59 (2C), 172.50, 172.60 (Cxe2x95x90O); nil/z (FAB) 598.2745 (MH+, C29H39N7O5P requires 596.2750).
L-Alanine (cyclobutyl) ester p-toluene sulfonate salt 
Prepared according to Standard Procedure 2, except using benzene as solvent: from L-alanine (1.0 g, 11 mmol),p-TSA monohydrate (2.35 g, 12 mmol), cyclobutanol (0.9 ml, 11 mmol) and benzene (65 ml). The p-toluene sulfonate salt was isolated as a white solid (1.73 g, 5.5 mmol, 49%). 6H (d4-CH3OH, 300 MHz) 1.51 (d, 3H, CH3-ala, J =7), 1.75 (m, 2H, CH2-cBu), 2.14 (m, 2H, CH2-cBu), 2.37 (m, 5H, CH2-c]u and CH3, p-TSA), 4.05 (q, 1H, CH-ala, J=7), 5.08 (m, 1H, CH-cBu), 7.24 (d, 2H, ArH, p-TSA), 7.70 (d, 2H, ArH, p-TSA); xcex4C (d4-CH3OH, 75 MHz) 14.57 (CH2-cBu), 16.58 (CH3-ala), 21.73 (CH3-pTSA), 31.38, 31.44 (CH2-cBu), 50.16 (CH-ala), 72.47 (CH-cBu), 127-35 (o-Ph, p-TSA), 130.23 (m-Ph, p-TSA), 142.13 (ipso-C-CH3, p-TSA), 143.89 (ipso-C-S, p-TSA), 170.71 (Cxe2x95x90O).
Phenyl(cyclobutoxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.75 ml, 15.0 mmol), dry triethylamine (1.4 ml, 10.0 mmol), L-alanine (cyclopentane methyl) ester p-toluene sulfonate salt 7a (1.58 g, 5.0 mmol) and dry DCM (65 ml total). The crude product was obtained as a clear, colourless oil (1.13 g, 71%)
xcex4P (CDCl3, 121 M)8.96, 9.33
The product was redissolved in dry THF (5 ml) and used as a 0.226 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-911-purine-9-yl)-2-cyclopentene-1-methanol O[pheny](cyclobutoxy-L-alaninyl)]phosphate [Cf 1775]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol) phenyl(cyclobutyl methoxy-L-alaninyl) phosphorochloridate 7b (2.95 ml of 0.226 g/ml solution, 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 3.5 hrs. The crude residue was purified by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a pale, yellow oil, which solidified to a cream solid after trituration and coevaporation with diethyl ether (0.238 g, 0.4 mmol, 60%).
xcex4P (CDCl3, 121 MHz) 3.89, 3.93; xcex4H (CDCl3, 300M ) 0.63 (m, 2H, CH2-cPr), 0.87 (m, 2H, CH2-cPr), 1.39 (m, 3H, CH3-ala), 1.65 (m, 2H, CH2-cBu), 1.81 (m, 1H, 61H,), 2.04 (m, 2H, CH2-cBu), 2.36 (m, 2H, CH2-cBu), 2.80 (m, 1H, 6xe2x80x2Hb), 3.03 (m, 1H, CH-cPr), 3.17 (m, 1H, 4xe2x80x2H), 3.97 (m, 2H, NH-ala and CH-ala), 4.18 (m, 2H, 5xe2x80x2H), 4.98 (m, 3H, NH2 42 and OCH), 5.55 (m, 1H, 11H), 5.91 (m, 1H, 31H), 6.01 (m, 1H, NH-cPr), 6.10 (m, 1H, 2xe2x80x2H), 7.25 (m, 5H, ArH), 7.51 (d, 1H, 8H); xcex4C (CDCl3, 75 Mz) 7.80 (CH2-cPr), 13.82 (CH2-cBu), 21.42 (CH3-ala), 22.06 (CH-cPr), 30.52-30.63 (CH2-cBu), 35.01 (6xe2x80x2C), 46.01, 46.12 (4xe2x80x2C), 50.50 (CH-ala), 59.26 (1xe2x80x2C), 69.30 (CH-cBu), 70.19 (5xe2x80x2C), 115.25 (5C), 120.53, 120.59 (p-Ph), 125.28 (o-Ph), 130.05 (m-Ph), 131.53 (3xe2x80x2C), 135.97 (8C), 136.73, 136.85 (2xe2x80x2C), 151.08-151.17 (6C and ipso-Ph), 156.71 (4C), 160.44 (2C), 173.33 (Cxe2x95x90O).
L-Alanine (cyclopentyl) ester p-toluene sulfonate salt 
Prepared according to Standard Procedure 2, except using benzene as solvent: from L-alanine (1.6 g, 18 mmol), p-TSA monohydrate (3.8 g, 20 mmol), cyclopentanol (1.6 ml, 18 mmol) and benzene (100 ml). The p-toluene sulfonate salt was isolated as a beige solid (2.81 g, 8.5 mmol, 47%).
xcex4H (d4-CH3OH, 300M ) 1.51 (d, 3H, CH3-ala, J=7), 1.71 (m, 6H, CH2-cPnt), 1.92 (m, 2H, CH2-cPnt), 2.39 (m. 5H, CH2-cBu and CH3,p-TSA), 4.04 (q, 1H, CH-ala, J=7), 5.28 (m, 1H, CH-cPnt), 7.26 (d, 2H, ArH, p-TSA), 7.73 (d, 2H, ATH, p-TSA); xcex4C (d4-CH3OH, 75 MHz) 16.59 (CH3-ala), 21.72 (CH3-pTSA), 24.97 (CH2-cPnt), 33.81, 33.97 (CH2-cPnt), 50.31 (CH-ala), 81.37 (CH-cPnt), 127.36 (o-Ph, p-TSA), 130.25 (m-Ph, p-TSA), 142.20 (ipso-C-CH3, p-TSA), 143.79 (ipso-C-S, p-TSA), 171.17 (Cxe2x95x90O).
Phenyl(cyclopentyloxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.9 ml, 6.0 10 mmol), dry triethylamine (1.7 ml, 12.0 mmol), L-alanine (cyclopentane methyl) ester p-toluene sulfonate salt 8a (1.98 g, 6.0 mmol) and dry DCM (100 ml total). The crude product was obtained as a clear, colourless oil (1.8 g, 91%).
xcex4P (CDCl3, 121 MHz) 9.01, 9.37
The product was redissolved in dry THF (5 ml) and used as a 0.361 g/ml solution. (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(cyclopentoxy-L-alaninyl)]phosphate [Cf 1776]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol) phenyl(cyclobutyl meth-oxy-L-alaninyl) phosphorochloridate 8b (1.93 ml of 0.361 g/ml solution, 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 3.5 hrs. The crude residue was purified twice by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to a white foam after trituration and coevaporation with diethyl ether (0.254 g, 0.4 mmol, 62%).
xcex4P (CDCl3, 121 MHz) 3.97, 3.98; SH (CDCl3, 300 MHz) 0.64 (m, 2H, CH2-cPr), 0.87 (m, 2H, CH2-cPr), 1.38 (m, 3H, CH3-ala), 1.67 (m, 7H, 3xc3x97CH2-cPent and 61H.), 1.86 (m, 2H, CH2-cPent), 2.81 (m, 1H, 6xe2x80x2Hb), 3.04 (m, 1H, CH-cPr), 3.18 (m, 1H, 4xe2x80x2H), 3.96 (m, 2H, Nil-ala and CH-ala), 4.21 (m, 2H, 5xe2x80x2H), 5.02 (bs, 1H, NH2), 5.18 (m, 1H, OCH), 5.56 (m, 1H, 1xe2x80x2H), 5.91 (m, 1H, 31H), 5.98 (bs, 1H, NH-cPr), 6.11 (m, 1H, 2xe2x80x2H), 7.25 (m, SH, ArH), 7.51 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 7.78 (CH2-cPr), 21.42, 21.48 (CH3-ala), 24.07 (CH-cPr), 32.91-(CH2-cPent), 33.05, 33.08 (6xe2x80x2C), 34.97, 35.02 (CH2-cPent), 46.02, 46.12 (4xe2x80x2C), 50.71 (CH-ala), 59.21, 59.25 (1xe2x80x2C), 69.22, 69.29 (5xe2x80x2C), 78.90 (OCH), 115.23 (5C), 120.55-120.61 (p-Ph), 125.28 (o-Ph), 130.05 (m-Ph), 131.53, 131.59 (3xe2x80x2C), 135.87, 135.97 (8C), 136.73,136.86 (2xe2x80x2C), 151.09, 151.18 (6C and ipso-Ph), 156.71 (4C), 160.44 (2C), 173.71, 173.80 (Cxe2x95x90O).
L-Alanine (phenethyl) ester p-toluene sulfonate salt 
Prepared according to Standard Procedure 2, from L-alanine (1.0 g, 11 mmol), p-TSA monohydrate (2.35 g, 12 mmol), phenethyl alcohol (1.3 ml, 11 mmol) and toluene (65 ml). The p-toluene sulfonate salt was isolated as an off-white solid (4.0 g, 10.9 mmol, 97%).
xcex4H (d4-CH3OH, 300 MHz) 1.46 (d, 3H, CH3-ala, J=7), 2.32 (2, 3H, CH3, p-TSA), 2.93 (t, 2H, CH2Ph, J=7), 4.07 (q, 1H, CH-ala, J=7), 4.37 (m, 2H, 0-CH2) 7.22 (m, 7H, ArH, p-TSA and PhH), 7.78 (d, 2H, ArH, p-TSA); xcex4C (d4-CH3OH, 75 MHz) 16.80 (CH3-ala), 22.06 (CH3-pTSA), 36.20 (CH2-Ph), 50.41 (CH-ala), 68.28 (O-CH2), 127.70,127.83 (o-Ar and o-Ph,p-TSA), 129.81 (p-Ar), 130.13, 130.48 (m-Ar and m-Ph, p-TSA), 139.23 (ipso-ArC), 142.30 (ipso-C-CH3, p-TSA), 143.83 (ipso-C-S, p-TSA), 171.44 (Cxe2x95x90O).
Pheny(phenethoxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.5 ml, 3.3 mmol), dry triethylamine (0.93 ml, 6.7 mmol), L-alanine (phenethyl) ester p-toluene sulfonate salt 9a (1.232 g, 3.3 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, colourless oil (1.16 g, 94%).
xcex4P (CDCl3, 121 MHz) 8.93, 9.25
The product was redissolved in dry THF (5 ml) and used as a 0.233 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(phenethoxy-L-alaninyl)]phosphate [Cf 1777]
Prepared according to Standard Procedure 4, from (1S,4R)-4(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol) phenyl(phenethoxy-L-alaninyl) phosphorochloridate 9b (3.3 ml of 0.233 g/ml solution, 2.1 mmol) and dry 1HF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 3 hrs. The crude residue was purified twice by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a pale, yellow oil, which solidified to a cream solid after trituration and coevaporation with diethyl ether (0.181 g, 0.3 mmol, 42%).
xcex4P (CDCl3, 121 MHz) 3.81, 3.86; xcex4H (CDCl3, 300M ) 0.65 (m, 2H, CH2CPr), 0.89 (m, 2H, CH2-cPr), 1.35 (m, 3H, CH3-ala), 1.71 (m, 1H, 61H,), 2.80 (m, 1H, 61Hb), 2.96 (m, 2H, CH2Ph), 3.03 (m, 1H, CH-cPr), 3.17 (m, 1H, 41H), 3.91 (m, 2H, NH-ala and CH-ala), 4.18 (m, 2H, OCH), 4.36 (m, 2H, 5xe2x80x2H), 4.99 (bs, 1H, NH2), 5.56 (m, 1H, 1xe2x80x2H), 5.92 (m, 2H, 3xe2x80x2H and NH-cPr), 6.09 (m, 1H, 21H), 7.26 (m, IOH, ArH and PhH), 7.52 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 6.36 (CH2-cPr), 19.98, 20.04 (CH3-ala), 22.65 (CHcPr), 33.46, 33.54 (6xe2x80x2C), 33.90 (CH2-Ph), 44.56, 44.66 (4xe2x80x2C), 49.21 (CH-ala), 57.79, 57.85 (1xe2x80x2C), 64.84 (OCH2), 67,82 (5xe2x80x2C), 113.79 (5C), 119.11-119.17 (p-Ph), 123.87 (p-Ar), 125.68 (o-Ph), 127.53 (o-Ar), 127.83 (m-Ph), 128.62 (m-Ar), 130.07, 130.14 (3xe2x80x2C), 134.48 (8C), 135.30, 135.39 (2xe2x80x2C), 136.25 (ipso-Ar), 149.63, 149.71 (6C and ipso-Ph), 155.25 (4C), 158.94 (2C), 172.75, 172.45 (Cxe2x95x90O).
L-Alanine (3-phenyl-1-propyl) ester p-toluene sulfonate salt 
Prepared according to Standard Procedure 2, from L-alanine (1.0 g, 11 mmol), p-TSA monohydrate (2.35 g, 12 mmol), 3-phenyl-1-propanol (1.5 ml, 11 mmol) and toluene (65 ml). Removal of the solvent gave the crude product as a yellow oil. Diethyl ether was added and the mixture was cooled for 30 mins. The resulting suspension was filtered to give the p-toluene sulfonate salt as a white solid (4.24 g, 11.2 mmol, 100%).
xcex4H (d4-CH3OH, 300 MHz) 1.53 (d, 3H, CH3-ala, J =7), 1.97 (m, 2H, CH2CH2Ph), 2.34 (2, 3H, CH3, p-TSA), 2.67 (t, 2H, CH2Ph, J=7), 4.10 (q, 1H, CH-ala, J =7), 4.20 (t, 2H, O-CH2, J=7) 7.22 (m, 7H, ArH, p-TSA and PhH), 7.75 (d, 2H, ArH, p-TSA); xcex4C (d4-CH3OH, 75 Ma) 16.77 (CH3-ala), 21.93 (CH3-pTSA), 31.63 (CH2CH2-Ph), 33.37 (CH2-Ph), 50.44 (CH-ala), 67.27 ()xe2x80x94CH2), 127.26-127.58 (o-Ar and o-Ph, p-TSA), 129.66-130.00 (p-Ar), 130.41 (m-Ar and m-Ph, p-TSA), 142.31 (ipso-ArC), 142.82 (ipso-C-CH3, p-TSA), 143.82 (ipso-C-S, p-TSA), 171.47 (Cxe2x95x90O).
Phenyl(phenethoxy-L-alaninyl)phosphorochloridate
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.5 ml, 3.3 mmol), dry triethylamine (0.93 ml, 6.7 mmol), L-alanine (3-phenyl-1-propyl) ester p-toluene sulfonate salt 9a (1.27 g, 3.3 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, pale brown oil (1.16 g, 90%).
xcex4P (CDCl3, 121 MHz) 8.94, 9.27
The product was redissolved in dry THF (5 ml) and used as a 0.231 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(3-phenyl-1-propoxy-L-alaninyl)]phosphate [Cf 1778]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol) phenyl(3-phenyl-1-propoxy-L-alaninyl) phosphorochloridate 10b (3.5 ml of 0.231 g/ml solution, 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 4 hrs. The crude residue was purified three times by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a pale, yellow oil, which solidified to an off-white foam after trituration and coevaporation with diethyl ether (0.330 g, 0.5 mmol, 75%). bp (CDCl3, 121 MHz) 3.89, 3.91; xcex4H (CDCl3, 300 MHz) 0.63 (m, 2H, CH2-cPr), 0.88 (m, 2H, CH2CPr), 1.42 (m, 3H, CH3-ala), 1.72 (m, 1H, 61Ha), 1.98 (CH2CH2Ph), 2.69 (CH2Ph), 2.80 (m, 1H, 6xe2x80x2Hb), 3.04 (m, 1H, CH-cPr), 3.18 (m, 1H, 4xe2x80x2H), 4.07 (m, 6H, NH-ala, CH-ala, OCH and 5xe2x80x2H), 5.00 (bs, 1H, NH2), 5.56 (m, 1xe2x80x2H, 1xe2x80x2H), 5.91 (m, 2H, 3xe2x80x2H and NH-cPr), 6.10 (m, 1H, 2xe2x80x2H), 7.25 (m, 10H, ArH and Phi), 7.52 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 6.35 (CH2-cPr), 20.06, 20.12 (CH3-ala), 22.65 (CH-cPr), 29.02 (CH2CH2Ph), 30.97 (CH2Ph), 33.48, 33.55 (6xe2x80x2C), 44.57, 44.67 (4xe2x80x2C), 49.26 (CH-ala), 57.78, 57.84 (1xe2x80x2C), 63.84 (OCH2), 67.88 (5xe2x80x2C), 113.83 (5C), 119.10, 119.15 (p-Ph and p-Ar), 123.86 (oPh), 125.09 (o-Ar), 127.33, 127.47 (m-Ph), 128.63 (m-Ar), 130.10, 130.17 (3xe2x80x2C), 134.47, 134.58 (8C), 135.27, 135.37 (2xe2x80x2C), 139.81 (ipso-Ar), 149.65, 149.74 (6C and ipso-Ph), 155.27 (4C), 158.98 (2C), 172.49, 172.58 (Cxe2x95x90O).
L-Alanine (4-phenyl-1-butyl) ester p-toluene sulfonate salt 
Prepared according to Standard Procedure 2, from L-alanine (1.0 g, 11 mmol), p-TSA monohydrate (2.35 g, 12 mmol), 4-phenyl-1-butanol (1.7 ml, 11 mmol) and toluene (65 ml). Removal of the solvent gave the crude product as a clear, colourless oil, which solidified to a white solid after trituration and coevaporation with petrol (60/80) (4.4 g, 11.2 mmol, 100%).
xcex4H (d4-CH3OH, 300 MHz) 1.55 (d, 3H, CH3-ala, J=7), 1.74 (m, 4H, xe2x80x94(CH2)2CH2Ph), 2.41 (2, 3H, CH3,p-TSA), 2.67 (m, 2H, CH2Ph), 4.12 (q, 1H, CH-ala, J=7), 4.28 (m, 2H, O-CH2) 7.25 (m, 7H, ArK, p-TSA and PhH), 7.75 (d, 2H, ArR, p-TSA); xcex4C (d4-CH3OH, 75 MHz) 16.65 (CH3-ala), 21.74 (CH3-pTSA), 29.18, 29.50 (OCH2(CH2)2CH2-Ph), 36.72 (CH2-Ph), 50.27 (CH-ala), 67.74 (O-CH2), 127.31, 127.36 (o-Ar and o-Ph,p-TSA), 129.79 (p-Ar), 130.25 (m-Ar and m-Ph, p-TSA), 142.14 (ipso-ArC), 143.64, 143.87 (ipso-C-CH3 and ipso-C-S, p-TSA), 171.47 (Cxe2x95x90O).
Pheny)(4-phenyl-1-butoxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.5 ml, 3.3 mmol), dry triethylamine (0.93 ml, 6.7 mmol), L-alanine (4-phenyl-1-butyl) ester p-toluene sulfonate salt 11a (1.32 g, 3.3 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, pale brown oil (1.13 g, 85%).
xcex4P (CDCl3, 121 MHz) 8.89, 9.24
The product was redissolved in dry THEF (5 ml) and used as a 0.226 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(4-phenyl-1-butoxy-L-alaninyl)]phosphate [Cf 11779]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol) phenyl(4-phenyl-1-butoxy-L-alaninyl) phosphorochloridate 11b (3.7 ml of 0.226 g/ml solution, 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 4 hrs. The crude residue was purified by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to an off-white foam after trituration and coevaporation with diethyl ether (0.314 g, 0.5 mmol, 69%).
xcex4P (CDCl3, 121 MHz) 3.87, 3.90; xcex4H (CDCl3, 300M ) 0.65 (m, 2H, CH2-cPr), 0.87 (m, 2H, CH2-cPr), 1.41 (m, 3H, CH3-ala), 1.71 (m, SH, (CH2)2CH2Ph and 61H,), 2.65 (m, 2H, CH2Ph), 2.80 (m, 1H, 6xe2x80x2Hb), 3.04 (m, 1H, CH-cPr), 3.17 (m, 1H, 41H), 4.06 (m, 6H, NH-ala, CH-ala, 5xe2x80x2H and OCH2xe2x80x94) 5.02 (bs, 1H, NH2), 5.56 (m, 1H, 11H), 5.90 (m, 11H, 3xe2x80x2H), 5.98 (bs, 1H, NH-cPr), 6.10 (m, 1H, 2xe2x80x2H), 7.25 (m, 10H, ArH and PhH), 7.52 (d, 1H, 8H); 8c (CDCl3, 75 MHz) 6.35 (CH2-cPr), 20.05. 20.11 (CH3-ala), 22.65 (CHcPr), 26.51 (CH2CH2CH2Ph), 27.02 (CH2(CH2)2Ph), 33.48, 33.55 (6xe2x80x2C), 34.32 (CH2Ph), 44.56, 44.67 (4xe2x80x2C), 49.22, 49.26 (CH-ala), 57.79, 57.83 (1xe2x80x2C), 64.40 (OCH2), 67.86, 67.94 (5xe2x80x2C), 113.75 (5C), 119.10, 119.15 (p-Ph and p-Ar), 123.85 (o-Ph), 124.88 (o-Ar), 127.33, 127.35 (m-Ph), 128.61 (m-Ar), 130.07, 130.12 (3xe2x80x2C), 134.44, 134.54 (8C), 135.30, 135.39 (2xe2x80x2C), 140.76 (ipso-Ar), 149.64-149.87 (6C and ipso-Ph), 155.26 (4C), 158.98 (2C), 172.53, 172.63 (Cxe2x95x90O).
L-Alanine (2-cyclobexyl ethyl) ester p-toluene sulfonate salt 
Prepared according to Standard Procedure 2, from L-alanine (1.0 g, 11 mmol), P-TSA monohydrate (2.35 g, 12 mmol), 2-cyclohexyl ethanol (1.56 ml, 11 mmol) and toluene (65 ml). Removal of the solvent gave the crude product as a clear, colourless oil, which solidified to a white solid after trituration and coevaporation with diethyl ether (2.8 g, 7.5 mmol, 67%).
xcex4H (d4-CH3OH, 300 M) 0.97 (m, 2H, CH2), 1.24 (m, 4H, 2xc3x97CH2), 1.42 (m, 1H, CH-cHx), 1.54 (d, 3H, CH3-ala, J =7), 1.63 (m, 2H, CHl], 1.75 (m, 4H, 2xc3x97CH2), 2.39 (s, 3H, CH3, p-TSA), 4.09 (q, 1H, CH-ala, J =7), 4.28 (m, 2H, O-CH2), 7.25 (d, 2H, ArH, p-TSA), 7.72 (d, 2H, ArH, p-TSA); xcex4C (d4-CH3OH, 75 MHz) 16.65 (CH3-ala), 21.74 (CH3-pTSA), 27.68 (CH2), 27.93 (CH2), 34.58 (CH2), 34.62 (CH2), 36.10 (CH-cHx), 50.27 (CH-ala), 66.05 [Oxe2x80x94CH2(CH2)2], 127.36 (o-Ph, p-TSA), 130.24 (m-Ph, p-TSA), 142.13 (ipso-C-CH3,p-TSA), 143.89 (ipso-C-S, p-TSA), 171.49 (Cxe2x95x90O).
Phenyl(2-cyclohexyl ethoxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.9 ml, 6.0 mmol), dry triethylamine (1.7 ml, 12.0 mmol), L-alanine (cyclohexyl ethyl) ester p-toluene sulfonate salt 12a (2.24 g, 6.0 mmol) and dry DCM (100 ml total). The crude product was obtained as a clear, colourless oil (1.86 g, 83%).
xcex4P (CDCl3, 121 MHz) 8.96, 9.31
The product was redissolved in dry IF (5 ml) and used as a 0.372 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(2-cyclohexyl-1-ethoxy-L-alaninyl)]phosphate [Cf 1780]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol) phenyl(cyclohexyl ethoxy-L-alaninyl) phosphorochloridate 12b (2.1 ml of 0.372 g/ml solution, 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 2.5 hrs. The crude residue was purified twice by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to an off-white foam after trituration and coevaporation with diethyl ether (0.302 g, 0.5 mmol, 69%).
xcex4P (CDCl3, 121 MHz) 3.91, 3.94; SH (CDCl3, 300 MHz) 0.64 (m, 2H, CH2-cPr), 0.91 (m, 4H, CH2 and CH2-cPr), 1.21 (m, 2H, CH2), 1.41 (m, 3H, CH3-ala), 1.52 (m, 2H, CH-cHx and 61H,), 1.70 (m, 6H, 3xc3x97CH2), 2.80 (m, 1H, 6xe2x80x2Hb), 3.04 (m, 1H, CH-cPr), 3.18 (m, 1H, 41H), 4.10 (m, 6H, NH-ala, CH-ala, OCH2 and 51H), 5.03 (bs, 1H, NH2), 5.56 (m, 1H, 11H), 5.96 (m, 1H, 31H), 5.98 (m, 1H, NH-cPr), 6.10 (m, 1H, 21H), 7.25 (m, 5H, Ar), 7.51 (d, 1H1, 8H); xcex4C (CDCl3, 75 MHz) 6-35 (Cl2-cPr), 20.05, 20.12 (CH3-ala), 22.69 (CH-cPr), 25.11 (CH2), 25.37 (CH2), 32.04, 32.07 (6xe2x80x2C), 33.45, 33.58 (CHcHx), 34.76 (CH2), 44.58, 44.69 (4xe2x80x2C), 49.28 (CH-ala), 57.78, 57.83 (1xe2x80x2C), 62.88 (OCH2), 67.86 (5xe2x80x2C), 113.82 (5C), 119.10-119.19 (p-Ph), 123.85 (o-Ph), 128.61 (m-Ph), 130.12 (3xe2x88x9dC), 134.44, 134.54 (8C), 135.28, 135.38 (2xe2x80x2C), 149.66-149.94 (6C and ipso-Ph), 155.28 (4C), 155.99 (2C), 172.57, 172.66 (Cxe2x95x90O).
L-Alanine (3-cyclobexyl-1-propyl) ester p-toluene sulfonate salt 
Prepared according to Standard Procedure 2, from L-alanine (1.0 g, 11 mmol), p-TSA monohydrate (2.35 g, 12 mmol), 3-cyclohexyl-1-propanol (1.7 ml, 11 mmol) and toluene (65 ml). The solvent was removed and diethyl ether was added. The resulting suspension was filtered to give the product as a white solid (3.9 g, 10.1 mmol, 90%).
xcex4H (d4-CH3OH, 300 MHz) 0.92 (m, 2H, CH2), 1.23 (m, 61, 3xc3x97CH2), 1.54 (d, 3H, CH3-ala, J=7), 1.71 (m, 7H, CH-cHx and 3xc3x97CH2), 2.39 (s, 3H, CH3, p-TSA), 4.10 (q, 1H, CH-ala, J =7), 4.22 (m, 2H, O-CH2), 7.25 (d, 2H, ArH, p-TSA), 7.72 (d, 2H, ArH, p-TSA); xcex4C (d4-CH3OH, 75 Mz) 16.66 (CH3-ala), 21.74 (CH3-pTSA), 27.36 (CH2), 27.83 (CH2), 28.11 (CH2), 34.80 (CH2), 34.90 (CH2), 39.03 (ICH-cHx), 50.27 (CH-ala), 68.27 (OCH2), 127.36 (o-Ph, p-TSA), 130.24 (m-Ph, p-TSA), 142.12 (ipso-C-CH3, p-TSA), 143.89 (ipso-C-S, p-TSA), 171.49 (Cxe2x95x90O).
Phenyl(3-cyclobexyl-I-propoxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.9 ml, 6.0 mmol), dry triethylamine (1.7 ml, 12.0 mmol), L-alanine (3-cyclohexyl-1-propyl) ester p-toluene sulfonate salt 13a (2.32 g, 6.0 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, pale yellow oil (2.31 g, 99%).
xcex4P (CDCl3, 121 MHz) 8.99, 9.35
The product was redissolved in dry THF (5 ml) and used as a 0.463 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(3-cyclobexyl-]-propoxy-L-alaninyl)]phosphate [Cf 1781]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol) phenyl(3-cyclohexyl-1-propoxy-L-alaniyl) phosphorochloridate 13b (1.8 ml of 0.463 g/ml solution, 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 2.5 hrs. The crude residue was purified by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to an off-white foam after trituration and coevaporation with diethyl ether (0.276 g, 0.4 mmol, 62%).
xcex4P (CDCl3, 121 MH) 3.89, 3.91; xcex4H (CDCl3, 300 Ma) 0.64 (m, 2H, CH2cPr), 0.89 (m, 2H, CH2-cPr), 1.21 (m, 6H, 3xc3x97CH2), 1.41 (in, 3H, CH3-ala), 1.66 (m, 8H, CH-cHx, 3xc3x97CH2 and 61Ha), 2.81 (m, 1H, 6xe2x80x2Hb), 3.04 (m, 1H, CH-cPr), 3.18 (in, 1H, 41H), 4.04 (m, 6H, NH-ala, CH-ala, OCH2 and 51H), 4.98 (bs, 1H, NH2), 5.56 (in, 1H, 11H), 5.91 (m, 1H, 31H and NHI-cPr), 6.11 (m, 1H, 21H), 7.26 (m, 5H, Ar), 7.57 (d, 1H, 8H); xcex4C (CDCl3, 75 M 6.37 (CH2-cPr), 20.09, 20.15 (CH3-ala), 22.66 (CH-cPr), 24.85 (CH2), 25.27 (CH2), 25.55 (CH2), 29.92 (CH2), 32.20, 32.33 (6xe2x80x2C), 33.49, 33.57 (CH-cHx), 36.22 (CH2), 44.58, 44.68 (4xe2x80x2C), 49.27 (CH-ala), 57.78, 57.83 (lxe2x80x2C), 65.01 (OCH2), 67.84 (5xe2x80x2C), 113.86 (SC), 119.10-119.19 p-Ph), 123.85 (o-Ph), 128.61 (m-Ph), 130.12 (3xe2x80x2C), 134.47, 134.57 (8C), 135.28, 135.38 (2xe2x80x2C), 149.65-149.74 (6C and ipso-Ph), 155.27 (4C), 158.96 (2C), 172.53, 172.64 (Cxe2x95x90O).
L-Alanine (4-cyclohexyl-1-butyl) ester p-toluene sulfonate salt 
Prepared according to Standard Procedure 2, from L-alanine (0.51 g, 5.8 mmol), p-TSA monohydrate (1.21 g, 6.3 mmol), 4-cyclohexyl-1-butanol (1.0 ml, 5.8 mmol) and toluene (65 ml). The p-toluene sulfonate salt was obtained as a white crystalline solid (2.15 g, 5.4 mmol, 93%).
xcex4H (d4-CH3OH, 300 MH) 0.92 (m, 2H, CH2), 1.17 (m, 6H, 3xc3x97CH2), 1.39 (m, 2H, Cl2), 1.54 (d, 3H, CH3-ala, J =7), 1.69 (m, 7H, CH-cHx and 3xc3x97CH2), 2.39 (s, 3H, CH3, p-TSA), 4.10 (q, 1H, CH-ala, J =7), 4.24 (m, 2H, O-CH2), 7.25 (d, 2H, ArH, p-TSA), 7.72 (d, 2H, ArH, p-TSA); 8c (d4-CH3OH, 75 MHz) 16.66 (CH3-ala), 21.74 (CH3-pTSA), 24.51 (CH2), 27.89 (CH2), 28.18 (CH2), 30.25 (CH2), 34.90 (CH2), 38.59 (CH2), 39.27 (CH-cHx), 50.27 (CH-ala), 67.94 (OCH2), 127.36 (o-Ph,p-TSA), 130.23 (m-Ph, p-TSA), 142.15 (ipso-C-CH3, p-TSA), 143.89 (ipso-C-S, p-TSA), 171.49 (Cxe2x95x90O).
Phenyl(4-cyclobexyl-1-butoxy-L-alaninyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.45 ml, 3.0 mmol), dry triethylamine (0.8 ml, 6.0 mmol), L-alanine (4-cyclohexyl-1-butyl) ester p-toluene sulfonate salt 14a (1.2 g, 3.0 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, brown oil (1.36 g,  greater than 100%).
xcex4P (CDCl3, 121 MHz) 8.91, 9.28
The product was redissolved in dry THF (5 ml) and used as a 0.272 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(4-cyclohexyl-1-butoxy-L-alaninyl)]phosphate [Cf 1782]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol 0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol) phenyl(3-cyclo-hexyl-1-propoxy-L-alaninyl) phosphorochloridate 14b (3.1 ml of 0.272 g/ml solution, 2.1 mmol) and dry THF (8 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 2.5 hrs. The crude residue was purified twice by column chromatography, using MeOH:CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to an off-white foam after trituration and coevaporation with diethyl ether (0.341 g, 0.5 mmol, 75%). 8p (CDCl3, 121 MHz) 3.89, 3.91; 8H (CDCl3, 300 MHz) 0.65 (m, 2H, CHz-cPr), 0.86 (m, 2H, CH2-cPr), 1.21 (m, 8H, 4xc3x97CH2), 1.41 (m, 3H, CH3-ala), 1.65 (m, 8H, CH-cHx, 3xc3x97CH2 and 61Hb), 2.81 (m, 1H, 6xe2x80x2Hb), 3.04 (m, 1H, CH-cPr), 3.19 (m, 1H, 4xe2x80x2H), 4.04 (m, 6H, NH-ala, CH-ala, OCH2 and 51H), 4.96 (bs, 1H, NH2), 5.56 (m, 1H, 1xe2x80x2H), 5.92 (m, 1H, 3xe2x80x2H and NH-cPr), 6.11 (m, 1H, 21H), 7.26 (m, 5H, Ar), 7.52 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 6.37 (CH2-cPr), 20.10, 20.16 (CH3-ala), 22.00 (CH2), 22.65 (CH-cPr), 25.34 (CH2), 25.64 (CH2), 27.77 (CH2), 32.28 (CH-cHx), 33.48, 33.56 (6xe2x80x2C), 36.45 (CH2), 44.58, 44.68 (4xe2x80x2C), 49.24 (CH-ala), 57.79, 57.84 (1xe2x80x2C), 64.69 (OCH2), 67.84, 67.94 (5xe2x80x2C), 113.86 (5C), 119.10-119.19 p-Ph), 123.86 (o-Ph), 128.62 (m-Ph), 130.11,130.17 (3xe2x80x2C), 134.47,134.57 (8C), 135.28, 135.39 (2xe2x80x2C), 149.65-149.74 (6C and ipso-Ph), 155.26 (4C), 158.96 (2C), 172.54, 172.64 (Cxe2x95x90O).
Phenyl(methoxy-L-valinyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.45 ml, 3.0 mmol), dry triethylamine (0.8 ml, 6.0 mmol), L-valine methyl ester hydrochloride salt (0.5 g, 3.0 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, colourless oil (0.922 g,  greater than 100%).
xcex4P (CDCl3, 121 MHz) 8.99, 9.37
The product was redissolved in dry TEF (5 ml) and used as a 0.184 g/ml solution.
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(methoxy-L-valinyl)phosphate [Cf 1686]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol), phenyl(methoxy valinyl)phosphorochloridate 15a (3.5 ml of 0.184 g/ml solution, 2.1 mmol) and dry THF (5 ml). The reaction mixture was stirred for 16 hrs, after which time a further 1.5 ml of the solution of 15a was added. The reaction mixture was stirred for a further 4 hrs. The crude residue was purified by column chromatography, using MeOH:DCM (5:95) as eluent, to give the product as a clear, colourless oil, which solidified to a white foam after trituration and coevaporation with diethyl ether (0.161 g, 0.3 mmol, 41%).
xcex4P (CDCl3, 121 MHz) 4.65, 4.74; BH (CDCl3, 300 MHz) 0.66 (m, 2H, CH2-cPr), 0.94 [m, 8H, CH2-cPr and CH(CH3)2], 1.71 (m, 1H, 61H.), 2.06 [m, 1H, CH(CH3)2], 2.81 (m, 1H, 6xe2x80x2Hb), 3.04 (m, 1H, CH-cPr), 3.18 (m, 1H, 41H), 3.52 (m, 1H, CH-val), 3.70 (d, 3H, OCH3), 3.83 (m, 1H, NH-val), 4.22 (m, 2H, 51H), 4.86 (bs, 2H, NH2), 5.56 (m, 1H, 1xe2x80x2H), 5.74 (bs, 1H, N1H-cPr), 5.93 (m, 1H, 31H), 6.11 (m, 1H, 21H), 7.27 (m, 5H, ArH), 7.52 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 6.37 (CH2-cPr), 16.36, 16.45 [CH(CH3)2], 22.65 (CH-cPr), 31.08, 31.16 [CH(CH3)2], 33.61 (6xe2x80x2C), 44.60, 44.70 (4xe2x80x2C), 51.05, 51.10 (OCH3), 57.78 (1xe2x80x2C), 58.96, 59.01 (CH-val), 67.90 (5xe2x80x2C), 113.91 (5C), 119.03-119.13 (o-Ph), 123.80 (p-Ph), 128.58 (m-Ph), 130.05, 130.14 (3xe2x80x2C), 134.47 (8C), 135.27, 135.39 (2xe2x80x2C), 149.66-149.84 (6C and ipso-Ph), 155.28 (4C), 158.96 (2C), 172.10, 172.19 (Cxe2x95x90O); m/z (FAB) 556.2428 (MH+, C26H35N7O5P requires 556.2437).
Phenyl(methoxy-L-leucinyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichiorophosphate (0.41 ml, 2.8 mmol), dry triethylamine (0.77 ml, 5.5 mmol), L-leucine methyl ester hydrochloride salt (0.5 g, 2.8 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, pale yellow oil (1.062 g,  greater than 100%).
xcex4P (CDCl3, 121 MHz)9.33, 9.51
The product was redissolved in dry THF (5 ml) and used as a 0.212 g/ml solution.
(1S, 4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(methoxy-L-leucinyl)phosphate [Cf1718]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-pun-9-yl)-2-cyclopentene-1-methanol 0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.4 ml, 1.4 mmol), phenyl (methoxy-L-leucinyl) phosphorochloridate 16a (3.2 ml of 0.212 g/ml solution, 2.1 mmol) and dry THF (10 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 2 hrs. The crude residue was purified twice by column chromatography, using MeOH: CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to a white foam after trituration and coevaporation with diethyl ether (0.211 g, 0.4 mmol, 53%).
xcex4P (CDCl3, 121 MHz) 3.98, 4.06; 8H (CDCl3, 300 MHz) 0.64 (m, 2H, CH2CPr), 0.89 [m, 8H, CH2-CPr and CH(CH3], 1.51 (m, 2H, CH2leu), 1.69 [(m, 2H, CH(CH3)2 and 6xe2x80x2Ha], 2.80 (m, 1H,.6xe2x80x2Hb), 3.04 (m, 1H, CH-cPr), 3.16 (m, 1H, 4xe2x80x2H), 3.67 (m, 1H, CH-leu), 3.69 (d, 3H, OCH3), 3.98 (m, 1H, NH-leu), 4.19 (m, 2H, 51H1), 4.97 (bs, 2H, NH2), 5.55 (m, 1H, 1xe2x80x2H), 5.91 (m, 1H, NH-cPr and 3xe2x80x2H), 6.09 (m, 1H, 2xe2x80x2H), 7.25 (m, 5H, ArH), 7.51 (d, 1H, 8H); xcex4C (CDCl3, 75 Mz) 6.37 (CH2Pr), 20.69, 20.82 (CH3-1eu), 22.69 (CH-cPr), 23.28, 23.41 [CH(CH3)2], 33.54 (6xe2x80x2C), 42.6042.81 (CH2-leu), 44.59,44.70 (4xe2x80x2C), 51.19, (OCH3), 52.07, 52.16 (CH-leu), 57.80 (Ixc2x0 C), 67.91, 67.98 (5xe2x80x2C), 113.88 (5C), 118.99-119.14 (o-Ph), 123.80 (p-Ph), 128.58 (m-Ph), 130.06, 130.14 (3xe2x80x2C), 134.53 (8C), 135.27, 135.34 (2xe2x80x2C), 149.68-149.76 (6C and ipso-Ph), 155.28 (4C), 158.97 (2C), 173.12, 173.23 (Cxe2x95x90O); m/z(FAB) 570.2610 (MH+, C27H37N7O5P requires 570.2594).
Phenyl(methoxy-L-prolinyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dicllorophosphate (0.54 ml, 3.6 mmol), dry triethylamine (1.0 ml, 7.2 mmol), L-proline methyl ester hydrochloride salt (0.6 g, 3.6 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, colourless oil (1.24 g,  greater than 100%).
xcex4P (CDCl3, 121 MHz) 9.02, 9.22
The product was redissolved in dry THF (5 ml) and used as a 0.248 g/ml solution.
(1S4R4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[pheny](methoxy-L-prolinyl)phosphate [Cf 1719]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.2 g, 0.7 mmol), xe2x80x2BuMgCl (1.0M in TBT: 1.4 ml, 1.4 mmol), phenyl(methoxy-L-prolinyl) phosphorochloridate 17a (2.6 ml of 0.248 g/ml solution, 2.1 mmol) and dry THF (10 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 20 hrs. The crude residue was purified twice by column chromatography, using MeOH: CHCl3 (4:96) as eluent, to give the product as a clear, colourless oil, which solidified to a white foam after trituration and coevaporation with diethyl ether (0.168 g, 0.3 mmol, 44%). Bp (CDCl3, 121 MHz) 2.83, 2.90; BH (CDCl3, 300M ) 0.65 (m, 2H, CH2-cPr), 0.90 (m, 2H, CH2-cPr), 1.92 (m, 5H, CH2CH2-pro and 6xe2x80x2Hb), 2.83 (m, 1H, 6xe2x80x2Hb), 3.04 (m, 1H, CH-cPr), 3.17 (m, 1H, 4xe2x80x2H), 3.45 (m, 2H, N-CH2-pro), 3.70 (d, 3H, OCH3), 4.13 (m, 1H, CH-pro), 4.30 (m, 2H, 51H), 4.87 (bs, 2H, NH2), 5.56 (m, 1H, l1H), 5.73 (s, 1H, NH-cPr), 5.91 (m, 1X, 31H), 6.12 (m, 1X, 2xe2x80x2H), 7.27 (m, 5H, ArH), 7.55 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 6.42 (CH2-cPr), 22.62 (CH-cPr), 23.91, 24.02 (CH2-pro), 30.38, 30.49 (CH2-pro), 33.54 (6xe2x80x2C), 44.61, 44.72 (4xe2x80x2C), 46.89 (N-CH2), 51.07, 51.19 (OCH3), 57.71, 57.80 (1xe2x80x2C), 58.84, 58.92 (CH-pro), 67.67, 67.75 (5xe2x80x2C), 113.87 (5C), 118.90-119.22 (o-Ph), 123.64, 123.73 (p-Ph), 128.55, 128.59 (m-Ph), 130.00 (3xe2x80x2C), 134.46 (8C), 135.42, 135.63 (2xe2x80x2C), 149.81, 149.90 (6C and ipso-Ph), 155.20 (4C), 158.87 (2C), 172.72, 173.23 (Cxe2x95x90O); m/z (FAB) 554.2283 (MH+, C26H33N7O5P requires 554.2281).
Phenyl(dibenzyloxy-L-aspartinyl)phosphorochloridate 
Prepared according to Standard Procedure 3, from phenyl dichlorophosphate (0.45 ml, 3.0 mmol), dry triethylamine (0.8 ml, 6.0 mmol), L-aspartate dibenzyl ester p-toluene sulfonate salt (1.46 g, 3.0 mmol) and dry DCM (60 ml total). The crude product was obtained as a clear, yellow oil (0.8024 g, 55%).
xcex4P (CDCl3, 121 M)9.43, 9.58
The product was redissolved in dry THF (5 ml) and used as a 0.16 g/ml solution.
(1S,4R)-4-(2-amino6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl(dibenzyloxy-L-aspartinyl)phosphate [Cf 1720]
Prepared according to Standard Procedure 4, from (1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol (0.157 g, 0.55 mmol), xe2x80x2BuMgCl (1.0M in THF: 1.1 ml, 1.1 mmol), phenyl(dibenzyloxy-1-aspartinyl) phosphorochloridate 18a (5.0 ml of 0.16 g/ml solution, 1.6 mmol) and dry THF (10 ml). TLC (8% MeOH in CHCl3) showed the reaction to be complete after 1.5 hrs. The crude residue was purified twice by column chromatography, using MeOH: CHCl3 (3:97) as eluent, to give the product as a clear, colourless oil, which solidified to an off-white foam after trituration and coevaporation with diethyl ether (0.284 g, 0.4 mmol, 70%). Sp (CDCl3, 121 ) 3.68, 4.24; SH (CDCl3, 300M ) 0.63 (in, 2H, CH2-cPr), 0.85 (m, 2H, CH2Pr), 1.63 (m, 1H, 61H.), 2.71 (m, 1H, 6xe2x80x2Hb), 3.06 (m, 2H, CH-cPr and 4xe2x80x2H), 4.14 (m, 2H, CH-asp, NH-asp), 4.34 (m, 2H, 5xe2x80x2H), 4.98 (bs, 2H, NH2), 5.06 (d, 2H, OCH2Ph), 5.13 (d, 2H, OCH2Ph), 5.53 (m, 1H, 1xe2x80x2H), 5.88 (in, 2H, NH-cPr and 31H), 6.01 (m, 1H, 2xe2x80x2H), 7.25 (m, 1H, ArH), 7.49 (d, 1H, 8H); xcex4C (CDCl3, 75 MHz) 6.35 (CH2-cPr), 22.64 (CH-cPr), 33.40 (6xe2x80x2C), 37.44, 37.60 (CH2-asp), 44.50, 44.57 (4xe2x80x2C), 50.20, 50.33 (CH-asp), 57.79 (1xe2x80x2C), 65.77 (OCH2Ph), 66.65 (OCH2Ph), 67.86, 68.00 (5xe2x80x2C), 113.85 (5C), 119.09-119.34 (o-Ph), 123.92 (p-Ph), 127.34-127.55 (m-Ph and nap-Bn), 128.61 (o-Bn), 130.06 (3xe2x80x2C), 134.00 (ipso-Bn), 134.03 (ipso-Bn), 134.61 (8C), 135.23, 135.27 (2xe2x80x2C), 149.47-149.91 (6C and ipso-Ph), 155.28 (4C), 158.95 (2C), 169.22, 169.43, 170.00, 17029 (Cxe2x95x90O).
(1S,4R)-4-(2-:amino-6-cyclopropylamino-9H-purin-9-yl-2-cyclopentene-1-methanol O-(phenyl (2-methylpropyl)oxyalaninyl phosphate) CF1672
This was prepared by Standard Procedure 4.70% yield.
xcex4P 3.87, 3.91.
xcex4H 0.64 (2H, m, CHaHb, CH1, Hb, cyclopropyl), 0.92 (8H, m, CHa, Hb, CHa, Hb, cyclopropyl, CH(CH3)2), 1.42 (3H, m, CH3 alaninyl), 1.71 (1H, m, 6xe2x80x2-HaHb), 1.92 (1H, m, CH(CH3)2), 2.81 (1H, m, 6xe2x80x2-HaHb), 3.03 (1H, m, CH2 cyclopropyl), 3.19 (1H, m, 4xe2x80x2-H), 3.87 (3H, m, CH alaninyl, CH2CH(CH3)2), 4.09 (1H, m, NH alaninyl), 4.20 (2H, m, 5xe2x80x2-H), 4.91 (2H, br s, NH2), 5.53 (1H, br m, 1xe2x80x2-H), 5.80 (1H, br s, NH-cyclopropyl), 5.92 (1H, m, 3xe2x80x2-H), 6.12 (1H, m, 2xe2x80x2-H), 7.31 (5H, in, Ph-H), 7.48 (1H, br d, 8H)
xcex4C 5.45 (CH2cyclopropylxc3x972), 17.01 (CH(CH3)2), 19.23, 19.29 (Me alaninyl), 21.74 (CH-cyclopropyl), 25.72 (CH(CH3)2), 32.58, 32.64 (6xe2x80x2-C), 43.66, 43.76 (4xe2x80x2-C), 48.35 (CH alaninyl), 56.90 (1xe2x80x2-C), 66.88, 66.96, 67.03 (5xe2x80x2-C), 69.57, 69.60 (CH2CH(CH3)2), 118.17, 118.20, 118.24, 118.27 (o-Ph, 5-C), 122.94 (p-Ph), 127.70 (m-Ph), 129.19, 129.24 (3xe2x80x2-C), 134.35, 134.45 (8-C, 2xe2x80x2-C), 148.81, 148.72 (i-Ph), 149.62, 149.76 (6-C), 154.34 (4-C), 158.91, 158.96 (2-C), 171.68, 171.58 (C(O) alaninyl).
MS m/e 570.2505 (M+, C27H36N7O5P requires 570.2515).
HPLC tR 33.11 min (0% CH3CN (0 min), 80% CH3CN (35 min), 80% CH3CN (45 min), 0% CH3CN (55 min)).
(1S ,4R)-442-amino6-cyclopropylamino-9H-purin-9-y)-2-cyclopentene-1-methanol O-(phenyl (2,2-dimethylpropyl)oxyalaninyl phosphate) CF1673
This was prepared by Standard Procedure 4.94% yield.
xcex4P 3.88, 3.94.
xcex4H 0.61 (2H, m, CHaHb, CHa,Hb, cyclopropyl), 0.85 (2H, br m, CHa,Hb, CHa, Hb, cyclopropyl), 0.91 (9H, s, C(CH3)3 ), 1.41 (3H, m, CH3 alaninyl), 1.70 (1H, m, 6xe2x80x2-HaHb), 2.78 (1H, m, 6xe2x80x2-HaHb), 3.03 (1H, m, CH cyclopropyl), 3.18 (1H, m, 4xe2x80x2-H), 3.81 (3H, m, CH alaninyl, CH2CH(CH3)2), 4.09 (1H, m, NH alaninyl), 4.20 (2H, m, 5xe2x80x2-H), 4.97 (2H, br s, NH2), 5.52 (1H, br m, 1xe2x80x2-H), 5.86 (1H, br s, NH-cyclopropyl, 3xe2x80x2-H), 6.08 (1H, m, 2xe2x80x2-H), 7.25 (5H, m, Ph-H), 7.48 (1H, br d, 8H).
xcex4C 7.89 (CH2-cyclopropylxc3x972), 21.74, 21.77 (Me alaninyl), 26.81 (C(CH3)3), 24.21 (CH-cyclopropy), 31.90 (C(CH3)3), 35.06 (6xe2x80x2-C), 46.10, 46.20 (4xe2x80x2-C), 50.79, 50.83 (CH alaninyl), 59-42 (1xe2x80x2-C), 66.35 (5xe2x80x2-C), 69.34, 69.41, 69.49 (CH2C(CH3)3), 116.41 (5-C), 120.62,120.66, 120.68, 120.72 (o-Ph), 125.39 (p-Ph), 130.15 (7n-Ph), 131.61, 131.65 (3xe2x80x2-C), 136.82, 136.90 (8-C, 2xe2x80x2-C), 151.16, 151.25 (6-C, i-Ph), 156.78 (4-C), 158.91, 160.44 (2-C), 174.09, 174.20 (C(O) alaninyl).
ES+m/e 584.2640 (MH+, C28H39N7O5P requires 584.2672).
HPLC tR 34.97 min (0% CH3CN (0 min), 80% CH3CN (35 min), 80% CH3CN (45 min), 0% CH3CN (55 min)).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-(phenyl (3-methylbutyl)oxyalaninyl phosphate) CF1674
This was prepared by Standard Procedure 4.47% yield.
xcex4P 3.87, 3.89:
xcex4H 0.57 (2H, m, CHaHb, CHa,Hb cyclopropyl), 0.80 (8H, m, CHaHb, CHa, Hb, cyclopropyl, CH(CH3)2 ), 1.30 (3H, m, CH3 alaninyl), 1.42 (2H, m, OCH2CH2), 1.62 (2H, m, 6xe2x80x2-HaHb, CH(CH3)2), 2.70 (1H, m, 6xe2x80x2-H3Hb), 2.92 (1H, br s, CH cyclopropyl), 3.07 (1H, m, 4xe2x80x2-H), 3.88 (3H, m, CH alaninyl, OCH2CH2), 4.07 (3H, m, NE alaninyl, 5xe2x80x2-H), 4.91 (2H, br s, NH2), 5.48 (1H, br m, 1xe2x80x2-H), 5.83 (2H, br s, NH-cyclopropyl, 3xe2x80x2-H), 6.03 (1H, m, 2xe2x80x2-H3, 7.18 (5H, m, Ph-H), 7.42 (1H, br d, 8H).
xcex4C 7.81 (CH2-cyclopropylxc3x972), 21.49, 21.56 (Me alaninyl), 22.79, 22.83 (CH(CH3)2), 24.10 (CH(CH3)2), 25.38 (CH-cyclopropyl), 34.91, 34.99 (OCH2CH2), 37.54 (6xe2x80x2-C), 46.01, 46.11 (4xe2x80x2-C), 50.70 (CH alaninyl), 59.25, 59.29 (1xe2x80x2-C), 64.63, 64.66 (OCH2CH2), 69.22, 69.30, 69.38 (5xe2x80x2-C), 116.17 (5-C), 120.53, 120.55, 120.59, 120.61 (o-Ph), 125.28 (p-Ph), 130.05 (m-Ph), 131.54, 131.60 (3xe2x80x2-C), 135.96 (8-C), 136.70, 136.81 (2xe2x80x2-C), 151.09, 151.17 (6-C, i-Ph), 156.68 (4-C), 160.34 (2-C), 173.94, 174.05 (C(O) alaninyl).
ES+m/e 584.2664 (MH+, C28H39N7O5P requires 584.2672).
HPLC tR 38.51 min (0% CH3CN (0 min), 80% CH3CN (35 min), 80% CH3CN (45 min), 0% CH3CN (55 min)).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9-H-purin-9-yl)2-cyclopentene-1-methanol O-(phenyl (cycloheptanyl)oxyalaninyl phosphate) CF1752
This was prepared by Standard Procedure 4.41% yield.
xcex4P 3.96, 3.98.
xcex4H 0.68 (2H, m, CHaHb, CHa,Hb, cyclopropyl), 0.99 (2H, m, CHa,Hb, CHa, Hb, cyclopropyl), 1.36 (5H, m, CH3 alaninyl, 5xe2x80x3-Ha,Hb,6xe2x80x3-HaHb), 1.80 (1 1H, m, 6xe2x80x2-Ha,Hb, 2xe2x80x3-H, 3xe2x80x3-H, 4xe2x80x3-H, 7xe2x80x3-H, 5xe2x80x3-HaHb, 6xe2x80x3-HaHb), 2.80 (1H, m, 6xe2x80x2-Ha,Hb), 3.12 (1H, br s, CH cyclopropyl), 3.22 (1H, m, 4xe2x80x2-H), 3.97 (2H, m, CH alaninyl, NH alaninyl), 4.20 (2H, m, 5xe2x80x2-H), 4.95 (1H, m, O-CH), 5.18 (2H, br s, NH2), 5.57 (1xe2x80x2H, br m, 1xe2x80x2-H), 5.90 (1H, m, 3xe2x80x2-H), 6.12 (1H, m, 2xe2x80x2-H), 6.25 (1xe2x80x2H, br s, NH cyclopropyl), 7.25 (SH, m, Ph-H), 7.51 (1H, br d, 8H).
xcex4C 15.08 (CH2-cyclopropylxc3x972), 28.76, 28.82 (Me alaninyl), 30.40, 30.44 (3xe2x80x3-C, 6xe2x80x3-C), 24.10 (CH(CH3)2), 31.57 (CH-cyclopropyl), 35.87 (4xe2x80x3-C, 5xe2x80x3-C), 41.26, 41.29, 41.31, 41.36 (6xe2x80x2-C), 42.24 (2xe2x80x3-C, 7xe2x80x3-C), 53.32, 53.42 (4xe2x80x2-C), 58.08 (CH alaninyl), 61.15 (1xe2x80x2-C), 66.62 (5xe2x80x2-C), 116.17 (5-C), 127.81, 127.85, 127.88, 127.91 (o-Ph), 132.54 (p-Ph), 137.32, 137.49 (m-Ph), 138.75 (3xe2x80x2-C), 143.21 (8-C), 144.13, 144.22 (2xe2x80x2-C), 158.40, 158.49 (6-C, i-Ph), 164.42 (4-C), 167.41 (2-C), 180.47, 180.51,180.59 (C(O) alaninyl).
ES+m/e 632.2719 (M[Na]+, C30H40N7O5Nap requires 632.2726).
HPLC tR 41.92 min (0% CH3CN (0 min), 80% CH3CN (35 min), 80% CH3CN (45 min), 0% CH3CN (55 min)).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopeutene-1-methanol O-(phenyl diethoxy-L-aspartyl phosphate) CF1714
This was prepared by Standard Procedure 4.54% yield.
xcex4P 3.76,4.19.
xcex4H 0.62 (2H, m, CHaHb, CHa,Hb, cyclopropyl), 0.88 (2H, m, CHaHb, CHa, Hb, cyclopropyl), 1.25 (6H, m, CH3xe2x80x94CH2xe2x80x94O aspartylxc3x972), 1.68 (1H, m, 6xe2x80x2-HaHb), 2.75 (2H, m, xe2x80x94(CO)xe2x80x94CHaHb, aspartyl, 61-HaHb), 2.97 (2H, m, CH cyclopropyl, xe2x80x94(CO)-CHaHb aspartyl), 3.16 (1H, m, 4xe2x80x2-H), 4.15 (8H, m, CH aspartyl, CH2xe2x80x94O aspartylxc3x972, NH aspartyl, 5xe2x80x2-H), 4.90 (2H, br s, NH2), 5.52 (1H, br m, 1xe2x80x2-H), 5.80 (1H, br s, NH-cyclopropyl), 5.90 (1H, m, 3xe2x80x2-H), 6.08 (1H, m, 2xe2x80x2-H), 7.21 (5H, m, Ph-H), 7.48 (1H, br d, 8H).
xcex4C 8.65 (CH2-cyclopropylxc3x972), 15.31 (CH3-CH2-O aspartylxc3x972), 24.92 (CH-cyclopropyl), 35.72 ((CO)-CH2 aspartyl), 39.74, 39.91 (6xe2x80x2-C), 46.82, 46.90 (4xe2x80x2-C), 52.41, 52.47 (CH aspartyl), 60.11 (1xe2x80x2-C), 62.22 (CH3xe2x80x94CH2xe2x80x94O(CO)CH2 aspartyl), 63.15 (CH3xe2x80x94CH2xe2x80x94C(CO)CH 15 aspartyl), 70.14, 70.27, 70.35 (5xe2x80x2-C), 116.12 (5-C), 121.33, 121.40, 121.49, 121.55 (o-Ph), 126.15 p-Ph), 130.86 (m-Ph), 132.36 (3xe2x80x2-C), 136.90 (8-C), 137.54 (2xe2x80x2-C), 151.81, 151.85 (6-C, i-Ph), 157.39 (4-C), 161-01 (2-C), 171.67, 171.81 (C(O)CH2 aspartyl), 172.38, 1172.48, 172.52, 172.62 (C(O) aspartyl).
ES+m/e 614.2393 (MH+, C28H37N7O7P requires 614.2492).
HPLC IR 30.37 mn (0% CH3CN (0 min), 80% CH3CN (35 min), 80% CH3CN (45 min), 0% CH3CN (55 min)).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-(phenyl methoxy-L-methionyl phosphate) CF1715
This was prepared by Standard Procedure 4.49% yield.
xcex4P 3.90, 4.03.
xcex4H 0.61 (2H, m, CHaHb, CHa, Hb, cyclopropyl), 0.86 (2H, m, CHaHb, CHa,Hb, cyclopropyl), 1.71 (1H, m, CH-CHaCHb methioninyl), 1.90 (1H, m, CH-Ca,CHb methioninyl), 2.01 (3H, d, CH3xe2x80x94Sxe2x80x94), 2.30 (1H, m, 6xe2x80x2-HaHb), 2.47 (2H, m, 5xe2x80x2-CH2), 2.78 (1xe2x80x2H, m, 6xe2x80x2-Ha,Hb), 2397 (1H, br m, CH cyclopropyl), 3.14 (1H, m, 4xe2x80x2-H), 3.70 (3H, d, CH3xe2x80x94Oxe2x80x94), 3.80 (1H, m, CH methioninyl) 4.17 (31H, m, NH methioninyl, 5xe2x80x2-H), 4.89 (2H, br s, NH2), 5.49 (1H, m1xe2x80x2-H), 5.80 (1H, br s, NH-cyclopropyl), 5.90 (1H, m, 31H), 6.08 (1H, ma, 2-H), 7.24 (5H, m, Ph-H), 7.43 (1H, br d, 8H).
xcex4C 6.52 (CH2-cyclopropylxc3x972), 14-42, 14.47 (CH3xe2x80x94Sxe2x88x92), 22.81 (CH-cyclopropyl), 28,63, 28.78 (S-CH2), 32.62, 32.73,32.81 (CH-CH2-methioninyl), 33.65 (67-C), 44.73,44.83 (4xe2x80x2-C), 51.67 (CH methioninyl), 57.99 (1xe2x80x2-C), 68.13, 68.20,68.27 (5xe2x80x2-C), 114.03 (5-C), 119.15, 119.22, 119.24, 119.30 (o-Ph), 124.05, 124.10 (p-Ph), 128.80 (m-Ph), 130.26, 130.30 (3xe2x80x2-C), 134.72 (8-C), 135.42, 135.47 (2xe2x80x2-C), 149.76, 149.80, 149.84, 149.89 (i-Ph), 150.08 (6-C), 155.38 (4-C), 159.06 (2-C), 172.25, 172.28, 172.32, 172.36 (C(O)).
ES+m/e 588.2053 (0% C26H34N7O5PS requires 588.2080).
HPLC tR 29.64 min (0% CH3CN (0 min), 80% CH3CN (35 min), 80% CH3CN (45 min), 0% CH3CN (55 min)).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2cyclopentene-1-methanol O-(phenyl methoxy-L-tryptophanyl phosphate) CF1750
This was prepared by Standard Procedure 4.70% yield.
xcex4P 3.88,4.01.
xcex4H 0.68 (2H, m, CHaHb, CHa,Hb cyclopropyl), 0.92 (2H, m, CHaHb, CHaxe2x80x2, Hbxe2x80x2cyclopropyl), 1.53 (1H, m, 6xe2x80x2-HaHb), 2.68 (1H, m, 6xe2x80x2-HaHb), 2.99 (2H, br m, CH cyclopropyl, 4xe2x80x2-H), 3.22 (2H, m, CH2-Trp), 3.66 (3H, d, CH3-O-), 3.93 (3H, m, NH Tip, 5xe2x80x2-H), 4.35 (1H, m, CH Trp), 4.94 (2H, br s, NH2), 5.49 (1H, m, 1xe2x80x2-H), 5.87 (2H, m, NH-cyclopropyl, 3xe2x80x2-H), 5.97 (1H, m, 2xe2x80x2-H), 7.01 (1H, m, 6xe2x80x3-H), 7.26 (7H, m, Ph-H, 4xe2x80x3-H, 5xe2x80x3-H), 7.46 (1H, m, 7xe2x80x3-H), 7.52 (1H, m, 2xe2x80x3-H), 8.63 (1H, br d, 8H).
xcex4C 7.81 (CH2-cyclopropylxc3x972), 24.10 (CH-cyclopropyl), 34.86, 34.91 (6xe2x80x2-C), 45.81, 45.90, 46.00 (4xe2x80x2-C), 52.97 (CH Trp), 59.24, 59.29 (1xe2x80x2-C), 69.14, 69.20 (5xe2x80x2-C), 109.86, 110.11 (3xe2x80x3-C), 111.72 (7xe2x80x3-C), 118.91 (5-C), 119.95, 120.04 (4xe2x80x3-C, 5xe2x80x3-C), 120.40, 120.47, 120.57, 120.63 (o-Ph), 122.49, 122.56 (6xe2x80x3-H), 123.70 (2xe2x80x3-C), 125.23, 125.29 (P-Ph), 127.79, 127.98 (9xe2x80x3-C), 130.04 (m-Ph), 131.27 (3xe2x80x2-C), 136.08 (8-C), 136.50, 136.55, 136.76, 136.87 (2xe2x80x2-C, 8xe2x80x3-C), 151.05, 151.14, 151.17, 151.26 (i-Ph, 6-C), 156.68 (4-C), 160.35 (2-C), 173.58, 173.66 (C(O)).
ES+m/e 643.2432 (MH+, C32H36N8O5P requires 643.2546).
HPLC tR 31.46 min (0% CH3CN (0 min), 80% CH3CN (35 mm), 80% CH3CN (45 ), 0% CH3CN (5 min)).
(1S,4R)-4-(2-amino-6-cyclpropylamino-9H purin-9-yl)-2-cyclopentene-1-methanol O-(phenyl methoxy-L-isoleucinyl phosphate) CF1751
This was prepared by Standard Procedure 4.60% yield. ps xcex4P 4.48, 4.54.
xcex40.68 (2H, m, CHaHb, CHa, Hb,cyclopropyl), 0.91 (8H, m, CHa, Hb, CHa,Hb, cyclopropyl, CH3xc3x972 isoleucinyl), 1.15 (1H, m, CHaCHb isoleucinyl), 1.45 (1H, m, CHaCHb isoleucinyl), 1.75 (2H, m, 6xe2x80x2-HaHb, CH3CH), 2.83 (]H, m, 6xe2x80x2-HaHb), 3.05 (1H, br m, CH cyclopropyl), 3.19 (1H, m, 4xe2x80x2-H), 3.62 (1H, m, NH isoleucinyl), 3.71 (3H, d, CH3xe2x80x94Oxe2x80x94), 3.88 (1H, m, CH isoleucinyl), 4.21 (2H, m, 5xe2x80x2-H), 4.91 (2H, br s, NH2), 5.55 (1H, m, 1xe2x80x2-H), 5.81 (1H, br s, NH-cyclopropyl), 5.93 (1H, m, 3xe2x80x2-H), 6.12 (1H, m, 2xe2x80x2-H), 7.28 (5H, m, Ph-H), 7.52 (1H, br d, 8H).
xcex4C 7.82 (CH2-cyclopropylxc3x972), 11-89 (CH3CH2), 15.72 (CH3CH), 24.08 (CH-cyclopropyl), 25.04, 25.13 (CH3CH2), 34.99 (6xe2x80x2-C), 39.49, 39.56, 39.64 (CH2CH), 46.04, 46.14 (4xe2x80x2-C), 52.46, 52.50 (CH isoleucinyl), 59.24, 59.44, 59.54 (1xe2x80x2-C), 69.34 (5S-C), 116.12 (5-C), 120.47, 120.53, 120.58 (o-Ph), 125.27 (p-Ph), 130.03 (m-Ph), 131.50, 131.57 (3xe2x80x2-C), 136.04 (8-C), 136.84, 136.74 (2xe2x80x2-C), 151.10, 151.18, 151.27 (i-Ph, 6-C), 156.69 (4-C), 161.06, 161.09, 161.35, 161.41 (2-C), 173.48, 173.53 (C(O)).
ES+m/e 570.2496 (MH+, C27H37N7O5P requires 570.2594).
HPLC IR 32.83, 33.14 min (0% CH3CN (0 min), 80% CH3CN (35 min), 80% CH3CN (45 min), 0% Cl3CN (55 min)).
(1S,4R)-4-(2-amino6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-(phenyl dimethoxy-L-glutamyl phosphate) CF1749
This was prepared by Standard Procedure 4.38% yield.
xcex4P 3.99.
xcex4H 0.68 (2H, m, CHaHb, CHa,Hb, cyclopropyl), 0.91 (2H, m, CHaHb, C CHa,Hb, cyclopropyl), 1.73 (1H, m, 6xe2x80x2-Ha, Hb), 2.12 (1H, m, C(O)CH2CHa,Hb), 2.38 (2H, m, C(O)CH2), 2.82 (1H, m, 6xe2x80x2-HaHb), 3.05 (1H, m, CH cyclopropyl), 3.18 (1H, n, 4xe2x80x2-H), 3.68 (3H, s, MeOC(O)CH2), 3.72 (3H, s, MeOC(O)CH), 3.85 (1H, m, NH glutyl), 4.10 (1H, m, CH glutyl), 4.21 (2H, m, 5xe2x80x2-H), 4.95 (2H, br s, NH2), 5.57 (1H, br m, 1xe2x80x2-), 5.88 (1H, br s, NH-cyclopropyl), 5.95 (1H, m, 3xe2x80x2-H), 6.10 (1H, m, 2xe2x80x2-H), 7.25 (5H, m, Ph-H), 7.54 (1H, br s, 8H).
xcex4C 7.82 (CH2-cyclopropylxc3x972), 24.12 (CH-cyclopropyl), 29.66, 29.73, 29.88 (C(O)CH2CH2), C(O)CH2CH2), 34.91 (6xe2x80x2-C), 46.02, 46.12 (4xe2x80x2-C), 52.19 (CH3OC(O)CH2CH2), 54.17, 54.28 (CH3OC(O)CH2), 54.17 (CH glutyl), 59.31 (1xe2x80x2-C), 69.50 (5xe2x80x2-C), 115.42 (5-C), 120.48, 120.51, 120.55, 120.58 (o-Ph), 125.39 (p-Ph), 130.09, 130.22 (m-Ph), 131.55, 131.60 (3xe2x80x2-C), 136.13 (8-C), 1136.68, 136.77 (2xe2x80x2-C), 150.98, 151.05, 151.13 (6-C), 151.76 (i-Ph), 156.65 (4-C), 160.99, 161.02, 161.08, 161.12 (2-C), 173.33, 173.43 (C(O)xc3x972 glutyl).
ES+m/e 600.2216 (MH+, C27H35N707P requires 600.2335).
HPLC IR 27.25 min (0% CH3CN (0 min), 80% CH3CN (35 min), 80% CH3CN (45 min), 0% CH3CN (55 min)).
(1S,4R)-4(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-(phenyl (methoxy-xcex1-ethyl-L-glycinyl phosphate) CF1783
This was prepared by Standard Procedure 4.44% yield.
xcex4P 4.10.
xcex4H 0.59 (2H, m, CHaHb, CHa, Hb, cyclopropyl), 0.83 (5H, br m, CHaHb, CHa,Hb, cyclopropyl, CH3-CH2), 1.68 (3H, m, CH3xe2x80x94CH2, 6xe2x80x2-H.), 2.69 (1H, m, 6xe2x80x2-Ha,Hb), 2.91 (1H, m, 4xe2x80x2-H), 3.06 (1H, m, CH cyclopropyl), 3.58 (3H, d, J 3.0, MeO), 3.90 (211, m, NH glycinyl, CH glycinyl), 4.07 (2H, m, 5xe2x80x2-H), 5.02 (2H, br s, NH2), 5.42 (1H, m, 1xe2x80x2-H), 5.75 (1H, m, 3xe2x80x2-H), 5.98 (1H, m, 2xe2x80x2-H), 6.03 (1H, m, NH cyclopropyl), 7.18 (5H, m, Ph-H), 7.41 (1H, br d, 8H).
xcex4C 7.76 (CH2-cyclopropylxc3x972), 9.68, 9.76 (CH3CH2), 24.12 (CH-cyclopropyl), 28.05 20 (CH3CH2), 35.01 (6xe2x80x2-C), 46.02, 46.13 (4xe2x80x2-C), 52.70, 52.73 (CH3O), 56.02 (1xe2x80x2-C), 59.25 (CH-ala), 69.38 (5xe2x80x2-C), 116.10 (5-C), 120.48, 120.50, 120.55, 120.57 (o-Ph), 125.27 (p-Ph), 130.04 (m-Ph), 131.51 (3xe2x80x2-C), 135.86 (8-C), 136.86 (2xe2x80x2-C), 151.08, 151.13, 151.22 (6-C, i-Ph), 156.67 (4-C), 160.40 (2-C), 173.84, 173.87, 173.92 (C(O) alaninyl).
ES+m/e 564.2094 (M[Na]+, C25H32N7O5NaP requires 564.2100).
HPLC tR 16.82, 16.84 min (0% CH3CN (0 min), 80% CH3CN (15 mm), 80% CH3CN (25 min), 0% CH3CN (35 min)).
(1S S4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-(phenyl (methoxy-a-phenyl(RS)glycinyl phosphate) CF1784
This was prepared by Standard Procedure 4.46% yield.
xcex4P 3.18,3.28,3.42,4.29.
Proton and Carbon NMR gave complex spectra, consistent with the racemised product.
ES+nm/e 612.2086 (M[Na+, C29H32N7O5NaP requires 612.2100).
HPLC tR 17.63, 18.50 min (0% CH3CN (0 min), 80% CH3CN (15 min), 80% CH3CN (25 min), 0% CH3CN (35 min)). (1:1.08 racemisation by HPLC)
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-(phenyl (methoxy-xcex1-butyl-L-glycinyl phosphate) CF1786
This was prepared by Standard Procedure 4.51% yield*.
Note: compound isolated as 6:1 (S:R) stereoisomeric mixture at the amino acid residue xcex1-carbon. Additional resonances in the 31P NMR spectra are noted at 4.35 and 5.18, corresponding to the minor configuration (R) amino acid residue containing diastereoisomers. 
xcex4P 4.10, 4.16.
xcex4H 0.51 (2H, m, CHa, Hb, CHa, Hb, cyclopropyl), 0.72 (5H, br m, CHaHb, CHa, Hb. cyclopropyl, CH3-CH2), 1.18 (4H, m, CH3-CH2-CH2-), 1.54 (3H, m, CH2-CH2-O, 6xe2x80x2-HaHb), 2.73 (1H, m, 6xe2x80x2-HaHb), 2.93 (111 m, 4xe2x80x2-H), 3.09 (]H, m, CH cyclopropyl), 3.52 (1H, m, CH glycinyl), 3.62 (3H, s, MeO), 3.87 (1H, m, NH glycinyl), 4.12 (2H, m, 5xe2x80x2-H), 4.75 (2H, br S, NH2), 5.45 (1H, m, 1xe2x80x2-H), 5.79 (2H, br s, NH-cyclopropyl, 3xe2x80x2-H), 6.00 (1H, m, 2xe2x80x2-H), 7.20 (5H, m, Ph-H), 7.42 (1H, br d, 8H).
xcex4C 7.76 (CH2-cyclopropylxc3x972), 14.23 (CH3CH2), 22.56 (CH3CH2), 24.14 (CH-cyclopropyl), 27.43, 27.50 (CH3CH2CH2), 34.50, 34.58 (CH2CH20), 35.01 (6xe2x80x2-C), 46.02, 46.12 (4-C), 52.66, 52.68 (CH3O), 54.87, 54.94 (1xe2x80x2-C), 59.20 (CH-ala), 69.30, 69.37 (5xe2x80x2-C), 115.18 (5-C), 120.29, 120.42, 120.50, 120.57 (o-Ph), 125.21 ip-Ph), 130.00 (m-Ph), 131.51, 131.54 (3xe2x80x2-C), 135.86 (8-C), 136.71, 136.76 (2xe2x80x2-C), 151.12, 151.16, 151.20, 151.25 (6-C, i-Ph), 156.73 (4-C), 160.49, 160.96 (2-C), 174.19, 174.26 (C(O) glycinyl).
ES+m/e 592.2428 (M[Na]+, C27H3N7O5NaP requires 592-2413).
HPLC IR 18.34, 18.41, min and 16.64 min (6:1) (0% CH3CN (0 min), 80% CH3CN (35 min), 80% CH3CN (45 min), 0% CH3CN (55 min)).
(1S,4R)-4-2-amino6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-(phenyl (methoxy-xcex1-propyl-L-glycinyl phosphate) CF1785
This was prepared by Standard Procedure 4.50% yield.
xcex4P 4.14,4.21.
xcex4H 0.62 (211, m, CHa,Hb, CHa,Hb, cyclopropyl), 0.86 (5H3, br m, CHaHb, CHa, Hb, cyclopropy], CH3xe2x80x94CH2), 1.32 (2H, m, CH3-CH2-), 1.63 (3H, m, CH3-CH2, 6xe2x80x2-Ha,Hb), 2.79 (1H, m, 6xe2x80x2-HaHb), 3.03 (1H, m, 4xe2x80x2-H, 3.18 (1H, m, CH cyclopropyl), 3.71 (3H, d, J 3.0, MeO), 3.97 (1H, m, CH glycinyl), 4.15 (3H, m, 5xe2x80x2-H, NH glycinyl), 5.09 (2H, br s, NH2), 5.55 (1H, m, 1xe2x80x2-H), 5.90 (1H, m, 3xe2x80x2-H), 6.08 (2H, m, 2xe2x80x2-H, NH cyclopropyl), 7.23 (5H, m, Ph-H), 7.52 (1H, br d, 8H).
xcex4C 7.55 (CH2-cyclopropylxc3x972), 13.98 (C1H3CH2), 18.62, 18.70 (CH3CH2), 24.15 (CH-cyclopropyl), 35.00 (6xe2x80x2-C), 36.89, 36.96 (CH2CH2O), 46.02, 46.12 (4xe2x80x2-C), 52.68 (CH3O), 54.70, 54.77 (1xe2x80x2-C), 59.21 (CH-ala), 69.31, 69.38 (5xe2x80x2-C), 115.24 (5-C), 120.45, 120.51; 120.57 (o-Ph), 125.22 (p-Ph), 130.01 (m-Ph), 131.54 (3xe2x80x2-C), 135.89 (8-C), 136.72, 136.78 (2xe2x80x2-C), 151.11, 151.16, 151.20, 15125 (6-C, i-Ph), 156.72 (4-C), 160.45, 160.95 (2-C), 174.18, 174.25 (C(O) alaninyl).
ES+m/e 578.2259 (M[Na]+, C26H34N7O5NaP requires 578.2257).
HPLC IR 17.57 ml (0% CH3CN (0 min), 80% CH3CN (35 mL), 80% CH3CN (45 min), 0% CH3CN (55 ml)).
(1S,4R)-4-(2-amino6cyclopropyl-2 -amino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-((P-(2 2-dimethoxy-L-propionic acid methyl ester)-phenyl)methoxyalaninyl phosphate) CF1671
This was prepared by Standard Procedure 4.24% yield.
xcex4P 3.72, 3.84.
xcex4H 0.56 (2H, m, CHa, Hb, CHa,Hb, cyclopropyl), 0.79 (2H, m, CHa,Hb, CHa, Hb, cyclopropyl), 1.30 (3H, m, CH3 alaninyl), 1.63 (1H, m, 6xe2x80x2-HaHb), 2.70 (1H, m, 6xe2x80x2-HaHb), 2.95 (1H, br s, 4xe2x80x2-H), 3.07 (3H, m, CH cyclopropyl, Ph-CH2-), 3.26 (6H, S, (OMe)2), 3.52 (3H, s, C(OMe)2COOMe), 3.61 (3H, s, COOMe alaninyl), 3.84-4.08 (4H, m, CH alaninyl, NH alaninyl, 5xe2x80x2-H), 4.99 (2H, br s, NH2), 5.46 (1H, br m, 1xe2x80x2-H), 5.81 (1H, br s, 3xe2x80x2-H), 6.02 (2H, m,3xe2x80x2-H, NH-cyclopropyl), 6.02 (1H, m, 2xe2x80x2-H), 7.02 (4H, m, Ph-H), 7.45 (1H, br d, 8H).
xcex4C 7.77 (CH2cyclopropylxc3x972), 21.37 (Me alaninyl), 24.01 (CH-cyclopropyl), 34.89 (6xe2x80x2-C), 39.55 (Ph-CH2), 45.97, 46.09 (4xe2x80x2-C), 50.53 ((MeO)2, CH3OO alaninyl), 52.65 (C(OMe)2COOMe), 59.28 (1xe2x80x2-C), 69.29 (5xe2x80x2-C), 103.27 (C(OMe)2),120.31, 120.38 (o-Ph), 122.94 p-Ph), 131.35 (m-Ph), 131.39 (3xe2x80x2-C), 136.79 (8-C, 2xe2x80x2-C), 150.14,150.05 (i-Ph, 6-C), 152.12 (4-C), 160.24 (2-C), 169.08 (C(OMe)2COOMC), 174.36, 174.46 (C(O) alaninyl).
ES+m/e 696.2531 ([M]+, C30H40N7O9NaP requires 696.2523).
HPLC IR 29.02 min (0% CH3CN (0 min), 80% CH3CN (35 min), 80% CH3CN (45 min), 0% CH3CN (55 min)).
(1xe2x80x2S,4R)-4-(2-amino-6cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-((p-methoxyphenyl)methoxyalaninyl phosphate) CF1815
This was prepared by Standard Procedure 4.23% yield.
xcex4P 4.23, 4.28.
xcex4H 0.72 (2H, m, CHaHb, CHa,Hb, cyclopropyl), 1.0 (2H, m, CHaHb, CHa, Hb cyclopropyl), 1.48 (3H, m, CH3 alaninyl), 1.82 (1H, in, 6-1HaHb), 2.80 (1H, m, 6xe2x80x2-HaHb), 3.11 (1H, br s, 4xe2x80x2-H), 3.25 (1H, m, CH cyclopropyl), 3.67 (1H, m, NH alaninyl), 3.77 (3H, s, COOMe alaninyl), 3.89 (3H, s, MeO-Ar), 4.14 (1H, m, CH alaninyl), 4.30 (2H, m, 5xe2x80x2-H), 4.94 (2H, br S, NH2), 5.65 (1H, br m, 1xe2x80x2-H), 5.83 (1H, br s, NH-cyclopropyl), 6.00 (1H, m, 3xe2x80x2-H), 6.17 (1H, m, 2xe2x80x2-H), 6.92 (2H, m, m-Ar), 7.23 (2H, m, o-Ar), 7.63 (1H, s, 8H).
xcex4C 7.81 (CH2-cyclopropylxc3x972), 21.46, 21.52 (Me alaninyl), 24.00 (CH-cyclopropyl), 34.96 (6xe2x80x2-C), 46.04, 46.14 (4xe2x80x2-C), 50.64 (CH300 alaninyl), 52.89 (CH-alaninyl), 56.02 (CH3O-Ar), 59.28 (1xe2x80x2-C), 69.30 (5xe2x80x2-C), 114.98 (m-Ph, 5-C), 121.42, 121.46, 121.52 (o-Ph), 131.52, 131.56 (3xe2x80x2-C), 135.98, (2xe2x80x2-C), 136.76, 136.87 (i-Ph), 144.61 (8-C), 156.71 (4-C), 157.01 (p-Ar), 161.40, 160.99 (2-C), 174.39, 174.50 (C(O) alaninyl).
HPLC IR 16.28 (0% CH3CN (0 min), 80% CH3CN (15 min), 80% CH3CN (25 min), 0% CH3CN (35 min)).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-((p-propoxyphenyl)methoxyalaninyl phosphate) CF1816
This was prepared by Standard Procedure 4.56% yield.
xcex4P 4.33, 4.41.
xcex4H 0.62 (2H, m, CHa,Hb, CHa,Hb, cyclopropyl), 0.82 (2H, m, CHaHb, CHa,Hb, cyclopropyl), 1.03 (311, t, J 6.0, CH3-CH2), 1.39 (311, m, CH3 alaninyl), 1.66 (1H, m, 6xe2x80x2-HaHb), 1.80 (2H, h, J 6.0, CH3-CH92), 2.79 (1H, m, 6xe2x80x2-HaHb), 3.01 (1H, br s, 4xe2x80x2-H), 3.12 (1H, m, CH cyclopropyl), 3.72 (3H, s, COOMe alaninyl), 3.89 (2H, t, J 6.0, CH2-O), 4.04 (2H1, m, CH alaninyl, NH alaninyl), 4.17 (211, m, 5xe2x80x2-H), 5.10 (21H, br s, NH2), 5.52 (1H, br m, 1xe2x80x2-H), 5.51 (1H, br s, NH-cyclopropyl), 5.89 (11H, m, 3xe2x80x2-H), 6.04 (1H, m, 2xe2x80x2-H), 6.81 (21 m, m-Ar), 7.11 (2H, m, o-Ar), 7.51 (1H, s, 8H).
xcex4C 7.77 (CH2-cyclopropylxc3x972), 10.91 (CH3-CH2), 21.39, 21.46 (Me alaninyl), 22.97 (CH3-CH2), 24.14 (CH-cyclopropyl), 34.96 (6xe2x80x2-C), 46.02, 46.13 (4xe2x80x2-C), 50.57, 50.65 (CH3OO alaninyl), 52.85, 52.87 (CH-alaninyl), 53.89 0 59.25 (1xe2x80x2-C), 69.16, 69.24, 69.33 (5xe2x80x2-C), 70.30 (CH2-O), 115.24,115.26 (5-C), 115.57 (m-Ph), 121.37, 121.40, 121.43, 121.46 (o-Ph), 131.51, 131.57 (3xe2x80x2-C), 135.93, (2xe2x80x2-C), 136.77, 136.85 (i-Ph), 144.47, 144.55 (8-C), 156.54, 156.73 (4-C), 160.45 (p-Ar), 160.91 (2-C), 174.48, 174.59 (C(O) alaninyl).
HPLC tR min (0% CH3CN (0 min), 80% CH3CN (15 min), 80% CH3CN (25 min), 0% CH3CN (35 min)).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[4-hydroxyacetophenone(methoxy-L-alaninyl)]-phosphoramidate Cf 1794
This was prepared by Standard procedure 4. The crude residue was purified twice by column chromatography, using MeOH:CHCl3 (3%:97) and MeOH:EtOAc (5:95) as eluent, to give the product as a white foam (30 mg, 17 mmol, 15%).
xcex4P 3.496.
xcex4SH 0.66 (m,2H,CH2-cPr), 0.85 (m,2H,CH2-cPr), 1.33 (m, 3H, CH3-CH), 1.7 (m, 1H, Hxe2x80x26), 2.53 (s, 3H, CH3-COPh), 2.8 (m, 1H, Hxe2x80x26), 2.9 (m, 1H, CH-cPr), 3.1 (m, 1H1, Hxe2x80x24), 3.6 (s, 3H, CH3-O), 3.9 (m, 1H, CH3-CH), 4.1 (m, 2H, Hxe2x80x25), 4.9 (m, 2H, NH2), 5.5 (m, 1H,xe2x80x21), 5.85 (m, 1H, Hxe2x80x23), 6.1 (m, 2H, Hxe2x80x22,NHcPr) 7.2 (dd, 2H, o-Ar), 7.5 (m, 1H, H8), 7.8 (dd, 2H, p-Ar)
xcex4C 6.371 (CH2cPr), 20 (CH-CH3aa), 21.671 (NHCH3), 25 (CH3CO), 33.458 (Cxe2x80x26), 44.55 (Cxe2x80x24),49.5 (CHaa), 51.4 (OCH3), 57.9 (Cxe2x80x21), 67.9 (Cxe2x80x25), 113.787 (C5), 120 (o-Ar), 122.22 (p-Ar), 128.743 (m-Ar), 130 (Cxe2x80x23), 134.53 (Cxe2x80x22), 135.31 (C8), 150.31 (i-Ar), 155.18 (C6), 156.342 (C2), 158.8 (C4), 173.004 (COOCH3), 198 (CO-Ar).
HPLC tr: 15.976 min (0% CH3CN (0 min), 80% CH3CN (15 min), 80% CH3CN (25 min), 0% CH3CN (35 min)).
(1S,4R)-4-(2-amino-6cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[4D-butylphenyl-(methoxy-L-alaninyl)-phosphate Cf 1795
The crude residue was purified twice by column chromatography, using MeOH:CHCl3 (3%:97) and MeOH:CH2Cl2 (5:95) as eluent, to give the product as a white foam (15 mg, 0.025 mmol, 4%),
xcex4P 3.934.00.
xcex4H 0.66 (m,2H,CH2-cPr), 0.85 (m,2H,CH2-cPr), 1.1 (m,3H, CH3-CH2),1.2 (m,4H, CH2-CH2), 1.33 (m, 3H, CH3-CH), 1.7 (m, 1H, Hxe2x80x26),2.5 (m, 2H, CH2-Ar), 2.8 (m, 1H, Hxe2x80x26),2.9 (m, 1H, CH-cPr), 3.1 (m, 1H, Hxe2x80x24),3.6 (s, 3H, CH3-O),3.9 (m, 1H, CH3xe2x80x94CH), 4.1 (m, 2, Hxe2x80x25),4.9 (m, 2H, NH2),5.5 (m, 1Hxe2x80x21), 5.85 (m, 1H, Hxe2x80x23), 6.1 (m, 2H, Hxe2x80x22,NHcPr), 7.2
xcex4C (dd, 211, o-Ar), 7.5 (m, 1H, H8),7.8 (dd, 2H,p-Ar). 6c 6.371 (CH2cPr), 14.345 (CH3xe2x80x94CH2), 21.49 (CH-CH3aa), 22.66 (CH2-CH3), 21.671 (NHCH3), 30.127 (CH2-CH2-CH12), 33.458 (Cxe2x80x26), 34.047.(CH2-Ar), 44.55 (Cxe2x80x24), 49.5 (CHaa), 51.4 (OCH3), 57.9 (Cxe2x80x21), 67.9 (Cxe2x80x25), 113.787 (C5), 120 (o-Ar), 122.22 (p-Ar), 128.743 (m-A), 130 (Cxe2x80x23),134.53 (C2),135.31 (C8),146.58 (i-Ax) 155.18 (C6),156.342 (C2),158.8 (C4),173.004 (COOCH)
HPLC tr: 19.591 mn (0% CH3CN (0 mm), 80% CH3CN (15 inn), 80% CH3CN (25 min), 0% CH3CN (35 min)).
(1S,4R)-4i(2-amino6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenylphenyl-(methoxy-L-alaninyl)]-phosphate Cf 1788
The crude residue was purified three times by column chromatography, using MeOH:CHCl3 (3:97) and MeOH:CH2Cl2 (5:95) and MeOH:AcOEt (3:97) as eluent, to give the product as a yellow foam (35 mg, 0.058 mmol, 8%).
xcex4P 3.94-3.96.
xcex4H 0.66 (m,2H,CH2-cPr), 0.85 (m,2H,CH2Pr), 1.33 (m, 3H, CH3xe2x80x94CH), 1.7 (m, 1H, Hxe2x80x26), 2.8 (m, 11H, Hxe2x80x26), 2.9 (m, 1H, CH-cPr), 3.25 (m, 1H, Hxe2x80x24), 3.6 (s, 3H, CH3-O), 4.1 (m, 1H, CH3-CH), 4.25 (m, 2H, Hxe2x80x25), 4.9 (m, 2H, N1H2), 5.5 (m, 1H,Hxe2x80x21), 5.85 (m, 1H, Hxe2x80x23), 6.15 (m, 2H, Hxe2x80x22,NHcPr), 7.35 (m, 9H, Ar), 7.6 (m, 1H, H8).
xcex4C 6.371 (CH2cPr), 21.49 (CH-CH3aa), 21.671 (NHCH3), 33.458 (Cxe2x80x26), 46.14 (C4), 50.671 (CHaa), 52.9 (OCH3),59.9 (Cxe2x80x21), 65.9 (Cxe2x80x25),115.787 (C5),120 (o-Ar), 122.22 (p-Ar), 128.743 (m-Ar), 130 (Cxe2x80x23), 134.53 (Cxe2x80x22), 135.31 (C8), 145.25 (i-Ar), 155.18 (C6), 156.342 (C2),158.8 (C4),173.004 (COOCH3).
HPLC tr: 19.147 min (0% CH3CN (0 mn), 80% CH3CN (15 min), 80% CH3CN (25 min), 0% CH3CN (35 min)).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenoxyphenyl-(methoxy-L-alaninyl)]-phosphate Cf 1787
The crude residue was purified twice by column chromatography, using MeOH:CHCl3 (3:97) and MeOH:CH2Cl2 (5:95) as eluent, to give the product as a yellow foam (35 mg, 0.058 mmol, 8%).
xcex4P 4.2124.184.
xcex4H 0.66 (m,2H,CH2-cPr), 0.85 (m,2H,CH2-cPr), 1.33 (m, 3H, CH3-CH), 1.7 (m, 1H, Hxe2x80x26), 2.8 (m, 1H, Hxe2x80x26), 2.9 (m, 1H, CH-cPr), 3.25 (m, 1H, H4), 3.6 (s, 3H, CH3-O), 4.1 (m, 1H, CH3-CH), 4.25 (m, 2H, Hxe2x80x25), 4.9 (m, 2H, NH2), 5.5 (m, 1H,Hxe2x80x21), 5.85 (m, 1xe2x80x2H, Hxe2x80x23), 6.15 (m, 2H, Hxe2x80x22,NHcPr), 7.35 (m, 9H, Ar), 7.6 (m, 1H, H8).
xcex4C 6.371 (CH2cPr), 21.49 (CH-CH3aa), 21.671 (NHCH3), 33.458 (Cxe2x80x26), 46.14 (Cxe2x80x24), 50.671 (CHaa), 52.9 (OCH3), 59.9 (Cxe2x80x21), 65.9 (Cxe2x80x25), 115.787 (CS), 120 (o-Ar), 122.22 p-Ar2), 128.743 (m-Ar), 130 (Cxe2x80x23), 134.53 (Cxe2x80x22), 135.31 (C8), 153.83 (i-Ar2, m-Ar1), 155.18 (C6), 156.342 (C2),158.8 (C4), 173.004 (COOCH3).
HPLC tr: 18.830 min (0% CH3CN (0 Min), 80% CH3CN (15 min), 80% CH3CN (25 min), 0% CH3CN (35 min))
(1S,4R)-4-(2-amino-6cyclopropy]amino9H-purin-9-yl)-2-cyclopentene-1-methanol O-phenyl methoxy-xcex1,xcex1-cyclopentylglycinyl]phosphate Cf1763
This was prepared by Standard procedure 4 in 77% yield.
31 p (CDCl3) 3.02, 3.09
1H (CDCl3) 0.56-0.61 (2H,m, CH2 (cpro)), 0.81-0.89 (2H, m, CH2 (cpro)), 1.58-1.78 (SH, m, CCH2CH2CH2CH2C, and H6xe2x80x2), 1.87-2.18 (4H, m, CCH2CH2CH2CH2C), 2.64-2.74 (1H, m, H6xe2x80x2), 2.83-3.09 (2H, m, CH(cpro), H4xe2x80x2), 3.60-3.62 (3H, s, OCH3(ala)), 4.044.19 (2H, m, H5xe2x80x2), 5.20 (2H, bs, NH2), 5.42-5.47 (1H, n, HIxe2x80x2), 5.77-5.83 (1H, m, H3xe2x80x2), 5.98-6.02 (1H, m, H2xe2x80x2), 6.20 (NH(cpro)), 7.06-7.27 (SH, m, Ar), 7.42-7.48 (1H, s, H8). 13C (CDCl3) 8.02 (CH2(cpro)), 24.37, 24.41 (CCH2CH2CH2CH2C), 34.73 (C6xe2x80x2), 38.48, 38.68,38.79, 38.87 (CCH2CH2CH2CH2C), 46.05, 46.15 (C4xe2x80x2), 52.99 (OCH3(ala)), 59.56, 59.60 (Clxe2x80x2), 67.16 (C (aa), 69.28, 69.37 (C5xe2x80x2), 114.76 (CS), 120.46, 120.52 (o-Ph), 125.22 (p-Ph),130.04 (m-Ph), 131.19 (C3xe2x80x2), 136.72 (C8), 137.13, 137.20 (C2xe2x80x2), 151.27, 151.31, 151.36,151.40 (C6), 155.56 (C4), 158.95 (C2),175.96, 176,00 (CO).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[phenyl methoxy-xcex1,xcex1-cylohexylglycinyl] phosphate Cf1764
This was prepared by Standard procedure 4 in 15% yield.
31 p (CDCl3) 2.89, 3.00
1H (CDCl3) 0.74 (2H,m, CH2 (cpro)), 1.01-1.03 (2H, m, CH2 (cpro)), 1.29-2.23 (11H, m, CCH2CH2CH2CH2CH2C, and H6xe2x80x2), 2.72-2.83 (1H, m, H6xe2x80x2), 3.17 (1H, bs, CH(cpro)), 3.35-3.43 (1H, m, H4xe2x80x2), 3.69-3.70 (3H, s, OCH3(aa)), 4.164.29 (2H, m, H5xe2x80x2), 5.52-5.66 (1H, m3 HIxe2x80x2), 5.79 (1H, bs, NH(cpro)), 5.85-5.90 (1H, m, H3xe2x80x2), 6.08-6.10 (1H, m, H2xe2x80x2), 7.15-7.35 (5H, m, Ar), 7.37-7.63 (1H, d, H8).
xe2x80x94C (CDCl3) 8.30 (CH2(cpro)), 21.49, 21.68, 21.84, 22.02, 22.11 (CCH2CH 2CH2CH2C), 25.14, 25.47, 25.72 (CCH2CH2CH2CH2CH2C ), 34.37, 34.59 (C6xe2x80x2), 46.06, 46.16 (C4xe2x80x2), 52.75, 53.12 (OCH3(aa)), 59.95, 60.19 (Clxe2x80x2), 69.22 (CSxe2x80x2), 120.41, 120.47 (o-Ph), 125.22 (p-Ph), 130.04 (m-Ph), 130.72, 130.82 (C3xe2x80x2), 137.41 (C8), 137.56 (C2xe2x80x2), 151.33, 151.43 (C6), 175.37 (CO).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentenel-methanol O-[phenyl methoxy-xcex1,xcex1-cylopropylglycinyl] phosphate Cf 1762
This was prepared by Standard procedure 4 in 69% yield.
31 P (CDCl3) 3.84
1H (CDCl3) 0.68(2H,m, CH2 (cpro)), 0.90-0.92 (2H, m, CH2 (cpro)), 1.16-1.49 (4H3 m, CCH2CH7C(aa)), 1.66-1.72 (1H, m, H6xe2x80x2), 2.72-2.82 (1H, m, H6xe2x80x2), 3.08-3.15 (2H, m, CH(cpro), H4xe2x80x2), 3.61-3.63 (3H, d, OCH3(aa)), 4.244.26 (2H, m, H5xe2x80x2), 5.24 (2H, bs, NH2), 5.53 (1H, bs, H1xe2x80x2), 5.87(1H, m, H3xe2x80x2), 6.07 (1H, m, H2xe2x80x2), 6.42-6.45 (1H, bs, NH(cpro)), 7.15-7.35 (SH, m, Ar), 7.56-7.61 (1H, d, HB).
13C (CDCl3) 7.92 (CH2(cpro)), 18.38 (CH2 (aa)), 35.20 (C6xe2x80x2), 52.88 (OCH3(aa)), 59.45 (Clxe2x80x2), 69.32 (C5xe2x80x2), 120.52 (o-Ph), 125.29 (p-Ph), 130.04 (m-Ph),137.03 (C2xe2x80x2), 151.13 (C6),160.96, 160.98 (C2), 174.35 (CO).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[p-(methoxycarbonyl)phenyl methboxy-L-alaninyl] phosphate Cf1766
This was prepared by Standard procedure 4 in 37% yield.
31P (CDCl3) 3.54, 3.58
1H (CDCl3) 0.66-0.69 (2H,m, CH2 (cpro)), 0.88-0.94 (2H, m, CH2 (cpro)), 1.38-1.43 (3H, t, CH3(ala)), 1.70-1.81 (1H, m, H6xe2x80x2), 2.76-2.89 (1H, m, H6xe2x80x2), 3.07 (1H, m, CH(cpro)), 3.21 (1H, mn, H4xe2x80x2), 3.71-3.73 (3H, d, OCH3(ala)), 3.94 (3H, s, COOCH3), 3.98-4.12 (1H, m, CH(ala)), 4.20-4.31 (2H, m, H5xe2x80x2), 5.19 (2H, bs, NH2), 5.54-5.57 (1H, m, H1xe2x80x2), 5.91-5.96 (1H, in, H3xe2x80x2), 6.09-6.14 (1H, m, H2xe2x80x2), 6.21 (1H, bs, NH(cpro)), 7.27-7.32 (2H, m, ArO,2H), 7.53-7.54 (1H, d, H8), 8.02-8.06 (2H, m, COAr,2H).
13C (CDCl3) 7.75 (CH2(cpro)), 21.22, 21.29, 21.46 (CH3(ala)),24.16 (NHCH), 34.83 (C6xe2x80x2), 45.97, 46.07 (C4xe2x80x2), 50.59 (CH(ala)), 52.57, 59.32 (OCH3 (ala), OCH3(Ph)), 59.27, 59.32 (Clxe2x80x2), 69.43 (C5xe2x80x2), 115.07, 115.11 (C5), 120.28, 120.31, 120.34, 120.38 (o-Ph), 127.07 (p-Ph), 131.58, 131.66 (m-Ph), 131.88 (C3xe2x80x2), 135.94, 136.04 (C2xe2x80x2), 136.61, 136.73 (C8), 151.31 (C6), 156.52 (C2), 160.97 (C4), 171.57 (CO), 174.30,174.39 (CO).
(1S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[p-(trifluoromethylthio)phenyl methoxy-L-alaninyl]phosphate Cf 1769
This was prepared by Standard procedure 4 in 34% yield.
31 P (CDCl3) 3.67, 3.88
1H (CDCl3) 0.81 (2H,bs, CH2 (cpro)), 1.06-1.08 (2H, m, CH2 (cpro)), 1.50-1.54 (3H, t, CH3(ala)), 1.83-1.93 (1I1, m, H6xe2x80x2), 2.87-2.99 (1H, m, H6xe2x80x2), 3.23-3.31 (1H, m, CH(cpro)), 3.82-3.84 (3H, d, OCH3(ala)), 4.1 44.15 (1H, mi, CH(ala)), 4.32-4.40 (2H, m, H5xe2x80x2), 5.65 (3H, bs, H1xe2x80x2, NH2), 6.01-6.04 (11, m, H3xe2x80x2), 6.19-6.23 (1H, m, H2xe2x80x2), 6.64 (1H, bs, NH(cpro)), 7.37-7.43(2H, m, Ar), 7.67 (1H, d, H8), 7.73-7.76 (2H, m, Ar). 7C (CDC)3) 8.16 (CH2(cpro)), 21.39, 21.45 (CH3(ala)), 34.45 (C6xe2x80x2), 46.09 (C4xe2x80x2), 50.66 (CH(ala)), 53.01 (OCH3 (ala)), 59.87 (Clxe2x80x2), 69.34 (C5xe2x80x2), 77.47, 77.67 (CF3S ?), 121.64, 121.69 (o-Ph), 127.07 (p-Ph), 136.99, 137.14 (C2xe2x80x2), 138.56 (C8), 153.36, 153.45 (C6), 160.93 (C4), 174.27 (CO).
(1S,4R)-4-(2-aminotcyclopropy]2 amino9H-purin-9-yl)2-cyclopentene-1-methano O-[p-(2-methoxyvinyl)phenyl methoxy-L-alaninyl] phosphate Cf 1767
This was prepared by Standard procedure 4 in 38% yield.
31p (CDCl3) 3.70, 3.74
1H (CDCl3) 0.58-0.61 (2H,bs, CH2 (cpro)), 0.81-0.85 (2H, m, CH2 (cpro)), 1.30-1.36 (3H, t, CH3(ala)), 1.61-1.72 (1H, m, H6xe2x80x2), 2.33 (31H, s, CH3CO), 2.70-2.79 (1H, m, H6xe2x80x2), 2.99 (1H, bs, CH(cpro)), 3.13(1H, bs, 1H4xe2x80x2), 3.64-3.65 (3H, d, OCH3(ala)), 3.924.01 (1H, m, CH(ala)), 4.114.21 (2H, m, H5xe2x80x2), 5.14 (3H, bs, H1xe2x80x2, NH2), 5.47-5.49 (1H, m, HIxe2x80x2), 5.82-5.87 (1H, m, H3xe2x80x2), 6.01-6.06 (1H, m, H12xe2x80x2), 6.12 (1H, bs, NH(cpro)), 6.57-6.63 (1H, dd, CH3COCHxe2x95x90CH), 7.14-7.46 (6H, m, H8, Ar, CH3COCHxe2x95x90).
13C (CDC]3) 7.95 (CH2(cpro)), 21.45 (CH3(ala)), 28.02 (CH3CO), 34.69 (C6xe2x80x2), 46.11 (C4xe2x80x2), 50.64 (CH(ala)), 52.99 (OCH3 (ala)), 59.53 (Clxe2x80x2), 121.03, 121.10, 121.17 (o-Ph), 127.39 (p-Ph), 130.13 (CH3COCHxe2x95x90CH), 131.44, 131.55 (C3xe2x80x2), 136.76 (C2xe2x80x2), 142.59 (CH3COCHxe2x95x90CH), 152.72 (C6),174.26,174.36 (CO(ala)), 198.70 (COCH3).
(1S,4R)-4-(2-amino-6cyclopropylamin o-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[p-(2-phenylcarbonylvinyl)phenyl methoxy-L-alaninyl] phosphate Cf 1771
This was prepared by Standard procedure 4 in 26% yield.
31p (CDCl3) 3.75,3.79
1H (CDCl3) 0.61-0.66 (2H,m, CH2 (cpro)), 0.85-0.91 (2H, m, CH2 (cpro)), 1.39-1.44 (3H, m, CH3(ala)), 1.67-1.86 (1H, m, H6xe2x80x2), 2.77-2.87 (1H, m, H6xe2x80x2), 3.04-3.05 (1H, bs, CH(cpro)), 3.19-3.21 (1H, bs, H4xe2x80x2), 3.72-3.73 (3H, d, OCH3(ala)), 4.024.13 (1H, m, CH(ala)), 4.194.29 (2H, m, H5xe2x80x2), 5.17 (3H, bs, H1xe2x80x2, NH2), 5.53-5.58 (1H, n, HIxe2x80x2), 5.90-5.95 (11H, m, H3xe2x80x2), 6.09-6.15 (2H, m, H12xe2x80x2, NH(cpro)), 7.24-8.08 (12H, m, Ar-, CH=C H, xe2x80x94Arxe2x80x94, H8).
13C (CDCl3) 7.85 (CH2(cpro)), 21.35, 21.41, 21.48 (CH3(ala)), 24.22 (CH(NH)), 34.80 (C6xe2x80x2), 46.01 (C4xe2x80x2), 50.67 (CH(ala)), 52.97 (OCH3 (ala)), 59.37 (Clxe2x80x2), 69.40 (C5xe2x80x2), 115.07 (C5), 121.01, 121.07, 121.14 (o-Ph), 128.92, 129.06 (p-Ph), 133.27 (C3xe2x80x2), 136.13, 136.23 (C2xe2x80x2), 138.53 (C8), 152.77, 152.86 (C6), 156.31 (C2), 160.97, 160.99 (C4), 174.31, 174.41 (CO(ala)), 190.76 (CO (Ar)).
(1S,4R)-4-(2-amino6-cyclopropylamin o-9H-purin-9-yl)-2-cyclopentene-1-methanol O[p-(2,2-dicyanovinyl)phenyl methoxy-L-alaninyl] phosphate Cf 1768
This was prepared by Standard procedure 4 in 10% yield.
31P (CDCl3) 4.54, 4.65
1H (CDCl3) 0.61-0.66 (2H,m, CH2 (cpro)), 0.85-0.91 (2H, m, CH2 (cpro)), 1.34-1.41 (3H, CH3(ala)), 1.67-1.83 (1H, m, H6xe2x80x2), 2.77-2.88 (1H, m, H6xe2x80x2), 2.95-2.97 (1H, m, H(cpro)), 3.23 (1H, bs, H4xe2x80x2), 3.68-3.70 (3H, d, OCH3(ala)), 3.99-4.03 (1H, m, CH(ala)), 4.22-4.32 (2H, m, H5xe2x80x2), 5.49-5.53 (11H, m, H1xe2x80x2), 5.99-6.03 (1H, m, H3xe2x80x2), 6.166.22 (1H, m, H2xe2x80x2), 6.94-6.97 (1H, dd, Ar-CH=CH), 7.36-7.41 (Ar), 7.64-7.65 (1H, d, H8), 7.92-8.16 (Ar).
13C (CDCl3) 6.56 (CH2(cpro)), 19.85 (CH3(ala)), 23.33 (CH(cpro)), 34.23 (C6xe2x80x2), 46.07 (C4xe2x80x2), 50.47, 50.53 (OCH3 (ala)), 51.78 (CH(ala)), 59.51 (Clxe2x80x2), 69.19, 69.29 (C5xe2x80x2), 113.84, 114.08 (C5), 121.14, 121.21, 121.27 (o-Ph), 128.49 (p-Ph), 130.74, 130.85 (m-Ph), 132.84 (C3xe2x80x2), 136.01 (C2xe2x80x2), 136.88, 136.99 (C8), 156.47 (C2), 160.99, 161.03 (C4), 174.27 (CO).
(1 S,4R)-4-(2-amino-6-cyclopropylamino-9H-purin-9-yl)-2-cyclopentene-1-methanol O-[o(carboxylate ethyl ester)phenyl methoxy-L-alaninyl]phosphate Cf1798
This was prepared by Standard procedure 4 in 24% yield.
31P (CDCl3) 4.03, 4.16
1H (CDCl3) 0.64-0.70 (2H,m, CH2 (cpro)), 0.92-0.93 (2H, d, CH2 (cpro)), 1.38-1.47 (6H, m, CH3(ala), CH3CH2O), 1.73-1.83 (1H, m, H6xe2x80x2), 2.78-3.24 (3H, m, H6xe2x80x2, H4xe2x80x2, CH(cyclo)), 3.64-3.72 (3H, s, OCH3(ala)), 4.084.20 (1H, m, CH(ala)), 4.234.45 (4H, m, H5xe2x80x2, CH2CH3), 5.21 (2H, bs, NH2), 5.55-560 (1H, m, H1xe2x80x2), 5.89-5.93 (1H, m, H3xe2x80x2), 6.13-6.18 (1H, m, H2xe2x80x2), 7.23-7.61 (1H, m, H8), 7.88-7.92 (1H, d, Ar).
31C (CDCl3) 7.95 (CH2(cpro)), 14.65 (CH3CH2), 21.33, 21.39, 21.68, 21.74 (CH3(ala)), 24.30 (NHCH), 34.80 (C6xe2x80x2), 46.04, 46.14 (C4xe2x80x2), 50.49 (CH(ala)), 52.74, 52.83 (OCH3 (ala)), 59.45 (Clxe2x80x2), 61.76, 61.82 (CH2CH3), 69.43, 69.51, 69.64 (C5xe2x80x2), 114.92 (C5), 122.93, 123.09, 123.60, 123.67125.26 (Ar), 131.34 (Ar), 131.77, 131.86 (C3xe2x80x2), 134.00 (Ar), 136.48 (C2xe2x80x2), 137.05 (C8), 150.20, 150.28 (C6), 155.88 (C2), 160.78, 160.86 (C4), 174.28, 174.39, 174.55, 174.65 (CO).